CMD–modified sensor chips

XanTec’s CMD sensor chips are based on a 2D (CMDP) or 3D hydrogel matrix consisting of carboxymethyl-dextran chains grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached through their amine, thiol, or aldehyde groups using established coupling chemistries such as EDC/NHS activation, thiol-maleimide coupling, or reductive amination. This versatility enables the immobilization of a wide spectrum of biomolecules including proteins, antibodies, peptides, nucleic acids, and small organic compounds.

The CMD sensor chip portfolio spans electrostatic immobilization capacities from a few thousand μRIU (CMDP) to about 50,000 μRIU (CMD700M), covering analytes from large viruses to small organic fragments. Owing to this versatility, CMD chips are used in biochemical research, assay development, quality control, trace analysis, and drug discovery. Different chain densities further enhance flexibility:

M variants (medium density, e.g., CMD200M, CMD700M) enable high ligand loading and elevated sensitivity for small-molecule kinetics and fragment screening.
L variants (low density, e.g., CMD50L, CMD200L) occupy less of the evanescent field, minimize diffusion effects, and are ideal for capture assays or large analytes.

Key features:

Versatile ligand coupling: Covalent coupling through amine, thiol, or aldehyde groups via standard chemistries (EDC/NHS, maleimide, reductive amination).
Wide immobilization range: From several thousand to ≈ 50,000 μRIU, suitable for analytes from whole cells and viruses to fragments < 300 Da.
Bioinert nanoarchitecture: Proprietary hydrophilic adhesion promoter combined with a hydrated CMD matrix minimizes nonspecific binding and matrix-analyte interactions.
Application versatility: Suitable for kinetic, equilibrium, and concentration analyses, as well as diverse screening applications in drug discovery.
High chemical stability: Withstands typical regeneration conditions, maintaining consistent response levels and kinetic behavior after multiple regeneration cycles.

Schematic illustration of a 2D CMDP (left) and 3D CMD (right) sensor chip. Red dots indicate the negatively charged carboxyl groups distributed along the green polysaccharide chains. The decaying red gradient represents the evanescent field.¹

Product code²	CMDP	CMD50L	CMD200L	CMD200M	CMD700M
Base coating	2D, ultra-short bioinert CM-dextran (high density)	3D, 50 nm bioinert CM-dextran (low density)	3D, 200 nm bioinert CM-dextran (low density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 700 nm bioinert CM-dextran (medium density)
Immobilization capacity [µRIU] ²	≈ 5,000	≈ 10,000	≈ 22,000	≈ 33,000	≈ 50,000
Recommended ligands	proteins peptides nucleic acids
Recommended analytes	proteins peptides nucleic acids viruses and cells	proteins peptides nucleic acids viruses and cells	small proteins peptides nucleic acids small molecules	small peptides small molecules fragments nucleic acids	small molecules fragments
Intended purpose	kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates applications requiring lowest diffusion limitation possible	similar to CM3 kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates applications requiring very low diffusion limitation minimizing nonspecific binding	kinetics of medium and small analytes equilibrium analysis applications requiring very low diffusion limitation immobilization of secondary antibodies and capture proteins	all-purpose, similar to CM5 kinetics of small analytes affinity and concentration analyses immobilization of capture proteins	similar to CM7 fragment-based drug discovery concentration analysis

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² This overview represents a selection of the full CMD sensor chip portfolio.

³ Preconcentration capacity determined by injecting 100 µg/mL bovine serum albumin (BSA) in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields can vary and depend strongly on the properties of the protein to be immobilized. Under optimal conditions, typical coupling efficiencies range from approximately 20–45% of the respective electrostatic preconcentration capacity, with acidic proteins generally exhibiting lower coupling efficiencies.

HC–modified sensor chips

XanTec’s HC sensor chips are based on a 2D (HCP) or 3D hydrogel matrix composed of highly flexible, bioinert polycarboxylate chains grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached through their amine, thiol, or aldehyde groups using established coupling chemistries such as EDC/NHS activation, thiol-maleimide coupling, or reductive amination. This versatility enables the immobilization of a wide range of biomolecules including proteins, antibodies, peptides, nucleic acids, carbohydrates, and small organic compounds.

The HC sensor chip portfolio spans electrostatic immobilization capacities from a few thousand μRIU (HCP) to ≈ 55,000 μRIU (HC1500M), covering analytes from large viruses to small organic fragments. The HC polycarboxylate matrix is more flexible and less bulky than carboxymethyl-dextran coatings, occupying a smaller fraction of the evanescent field. Its strongly hydrated polymer brush makes the surface highly bioinert and minimizes nonspecific binding. This combination provides high immobilization capacity with excellent diffusion properties, making HC chips a preferred alternative to CMD-based sensor surfaces in biochemical research, assay development, quality control, trace analysis, and drug discovery.

Key features:

Versatile ligand coupling: Covalent coupling through amine, thiol, or aldehyde groups via standard chemistries (EDC/NHS, maleimide, reductive amination).
Wide immobilization range: From several thousand to ≈ 55,000 μRIU, suitable for analytes from whole cells and viruses to fragments < 300 Da.
Bioinert nanoarchitecture: Proprietary hydrophilic adhesion promoter combined with a strongly hydrated polycarboxylate matrix minimizes nonspecific binding.
Application versatility: Suitable for kinetic, equilibrium, and concentration analyses, as well as diverse screening applications in drug discovery.
High chemical stability: Withstands typical regeneration conditions, maintaining consistent response levels and kinetic behavior after multiple regeneration cycles.
No polysaccharide backbone: Lack of carbohydrate motifs prevents unwanted interactions with lectins or carbohydrate-binding proteins, making HC sensor chips ideal for analyzing this class of biomolecules and bioelectrical impedance analysis (BIA) of carbohydrates.

Schematic illustration of a 2D HCP (left) and a 3D HC (right) sensor chip. Red dots indicate the negatively charged carboxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field.¹

Product code²	HCP	HC30M	HC200M	HC1500M
Base coating	2D, ultra-short bioinert polycarboxylate (high density)	3D, 30 nm bioinert polycarboxylate (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)	3D, 1500 nm bioinert polycarboxylate (medium density)
Immobilization capacity [µRIU]³	≈ 5,000	≈ 19,000	≈ 33,000	≈ 55,000
Recommended ligands	proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids viruses and cells carbohydrates	proteins peptides nucleic acids small molecules carbohydrates	peptides nucleic acids small molecules carbohydrates	small molecules fragments short nucleic acids small carbohydrates
Intended purpose	kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates applications requiring lowest diffusion limitation possible	all-purpose kinetics of small, medium, and large analytes especially suitable for weak binders with fast on- and off-rates applications requiring very low diffusion limitation immobilization of secondary antibodies and capture proteins	all-purpose kinetics of medium and small analytes equilibrium analysis concentration analysis immobilization of secondary antibodies and capture proteins	kinetics of small analytes fragment-based drug discovery affinity and concentration analyses immobilization of capture proteins

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² This overview represents a selection of the full HC sensor chip portfolio.

NeutrAvidin–modified sensor chips

XanTec NeutrAvidin-modified sensor chips are coated with a bioinert, charge-reduced (poly)carboxylate matrix (HLC), pre-functionalized with a recombinant 52 kDa NeutrAvidin tetramer. This immobilized NeutrAvidin efficiently captures biotinylated biomolecules—including proteins, nucleic acids, peptides, and other biotin-tagged ligands—under physiological conditions.

NeutrAvidin-modified sensor chips are especially tailored to reduce nonspecific binding in SPR assays. This is achieved by combining the deglycosylated, nearly charge-neutral (under physiological conditions) NeutrAvidin, with the highly bioinert, charge-reduced HLC coating, where most negative charges are neutralized. This unique combination renders the NAHLC sensor chips particularly suitable for assessing demanding analytes with high positive net charge.

Due to the exceptionally high binding affinity of NeutrAvidin for biotin (dissociation equilibrium constant ≈ 10⁻¹⁵ M), XanTec’s NAHLC sensor chips allow stable immobilization of biotinylated ligands with negligible dissociation.

Key features:

Reduced nonspecific binding: NAHLC is an alternative to streptavidin-modified sensor chips in assays affected by nonspecific binding.
Fast assay development: No preconcentration or surface activation required. Controlled ligand immobilization is performed under physiological conditions, reducing preparation time and ensuring reproducibility.
Exceptional stability: Exceptionally strong NeutrAvidin-biotin binding makes this surface compatible with most common regeneration protocols.
Oriented immobilization: Controlled biotinylation via biomolecular tools such as AviTag allows for maximum ligand activity and reproducible immobilization results.
Broad application range: Ideal for immobilization of a wide variety of biotinylated ligands, making it suitable for protein interaction studies, nucleic acid hybridization, antibody screening, and other biosensing applications.
Versatile capture capacity: Available on various HLC base coatings, offering different capture capacities to match specific experimental needs.

Schematic illustration of a 3D NAHLC sensor chip. Red and grey dots represent negatively charged carboxyl groups and charge-neutral hydroxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows immobilized NeutrAvidin (dark blue) binding a biotinylated protein ligand (yellow).¹

Product code	NAHCP²	NAHLC30M	NAHLC200M	NAHLC1500M
Base coating	2D, ultra-short bioinert polycarboxylate (medium density)	3D, 30 nm bioinert polycarboxylate (medium density, reduced charge)	3D, 200 nm bioinert polycarboxylate (medium density, reduced charge)	3D, 1500 nm bioinert polycarboxylate (medium density, reduced charge)
Capture immobilization capacity [µRIU]³	≈ 800	≈ 1,700	≈ 3,300	≈ 4,000
Recommended ligands	biotinylated proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids carbohydrates viruses and cells	proteins peptides nucleic acids carbohydrates	peptides nucleic acids carbohydrates	peptides nucleic acids carbohydrates small molecules
Intended purpose	kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates including carbohydrates capture immobilization of biotinylated secondary antibodies reduction of nonspecific binding	kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates including carbohydrates capture immobilization of biotinylated secondary antibodies reduction of nonspecific binding	all-purpose kinetics of medium analytes including carbohydrates especially suitable for situations requiring higher capture densities reduction of nonspecific binding	kinetics of medium and small analytes including carbohydrates especially suitable for situations requiring higher capture densities reduction of nonspecific binding

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² NAHCP is the only variant that employs the classic HC matrix instead of HLC.

³ Based on specific binding of 100 µg/mL biotinylated bovine serum albumin (BSA) in phosphate-buffered saline (PBS), with 1 µRIU corresponding approximately to 1 RU.

Azide–modified sensor chips

XanTec’s AZ sensor chips are coated with a bioinert polycarboxylate matrix (CMD or HC), pre-functionalized with low-molecular-weight azido groups grafted onto a hydrophilic adhesion promoter on a gold support. Ligands containing a compatible Click partner—such as dibenzocyclooctyne (DBCO)—can be covalently attached in a fast and selective manner via strain-promoted azide-alkyne cycloaddition (Click reaction).

Unlike classical EDC/NHS chemistry, DBCO/azide coupling is not time-critical; both reaction partners remain stable for weeks across pH 4—10. Their high selectivity minimizes side reactions and eliminates the need for quenching steps¹. Consequently, immobilization via Click coupling is reliable, convenient, and flexible. Owing to rapid reaction kinetics, coupling under physiological conditions is feasible, although maximum immobilization densities may be somewhat lower; thus, electrostatic preconcentration remains advantageous for achieving high ligand loading.

The AZ sensor chip portfolio spans electrostatic immobilization capacities from approximately 6000 μRIU (AZD50L) to 40,000 μRIU (AZHC1500M), covering analytes from large viruses to small organic fragments. The surface-bound azido groups are highly hydrophilic and possess a strong dipole moment, further enhancing the overall bioinertness of the coating.

Key features:

Versatile ligand coupling: Highly efficient and selective Click reaction enables covalent attachment of DBCO-functionalized ligands. Unlike classic amine coupling, ligand density can be iteratively increased after initial coupling, allowing fast optimization of immobilization levels and efficient sensor chip use.
Superior efficiency: The high efficiency of the Click reaction permits ligand immobilization even under physiological conditions.
DBCO functionalization: Ligands can be conveniently DBCO-modified using standard labeling protocols similar to biotinylation; site-specific conjugation via sortase-mediated ligation is also supported.
Wide immobilization range: From several thousand to ~40,000 μRIU, suitable for analytes from whole cells and viruses to fragments < 300 Da.
Bioinert nanoarchitecture: Combining a hydrophilic adhesion promoter, hydrated polycarboxylate matrix (CMD or HC), and surface azido groups minimizes nonspecific binding.
Application versatility: Suitable for kinetic, equilibrium, and concentration analyses, as well as diverse screening applications in drug discovery.

Schematic illustration of a 3D AZHC sensor chip. Short purple lines and red dots represent azido groups and negatively charged carboxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows a DBCO-modified antibody covalently immobilized via Click chemistry to azido functionalities.²

Product code³	AZD200M	AZHC30M	AZHC200M	AZHC1500M
Base coating	3D, 200 nm bioinert CM-dextran (medium density)	3D, 30 nm bioinert polycarboxylate (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)	3D, 1500 nm bioinert polycarboxylate (medium density)
Covalent immobilization capacity [µRIU]⁴	≈ 13,000 (30,000)	≈ 10,000 (18,000)	≈ 13,000 (26,000)	≈ 20,000 (40,000)
Recommended ligands	proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids	proteins peptides nucleic acids small molecules carbohydrates	peptides nucleic acids small molecules carbohydrates	small molecules fragments nucleic acids carbohydrates
Intended purpose	immobilization of DBCO-functionalized ligands kinetics of small to large analytes equilibrium analysis concentration analysis immobilization of secondary antibodies and capture proteins	immobilization of DBCO-functionalized ligands kinetic analysis of small to large analytes especially suitable for weak binders with fast association and dissociation rates Suitable for assays requiring very low diffusion limitation immobilization of secondary antibodies and capture proteins	immobilization of DBCO-functionalized ligands kinetics of small and medium analytes equilibrium analysis concentration measurements immobilization of secondary antibodies and capture proteins	immobilization of DBCO-functionalized ligands kinetics of small analytes fragment-based drug discovery affinity and concentration analyses immobilization of capture proteins

¹ This is valid only when the analyte lacks its own Click-reactive functionality; otherwise, unintended covalent attachment may occur.

² All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

³ Table includes a selection from XanTec’s full AZ sensor chip portfolio.

⁴ Covalent immobilization capacity was assessed by injecting 100 µg/mL DBCO-modified streptavidin in 5 mM sodium acetate buffer (pH 5.0), with 1 µRIU corresponding approximately to 1 RU. Electrostatic preconcentration capacity is shown in brackets.

DBCO–modified sensor chips

XanTec’s DBCO sensor chips (DC) are coated with a dibenzocyclooctyne (DBCO)-derivatized bioinert carboxymethyl-dextran (CMD) matrix grafted onto a hydrophilic adhesion promoter on a gold support. Ligands containing a compatible Click partner—such as an azide group—can be covalently attached rapidly and selectively via strain-promoted azide-alkyne cycloaddition (SPAAC, Click reaction).

Unlike classical EDC/NHS amine-coupling, DBCO/azide Click chemistry is not time-critical; both reaction partners remain stable for weeks across pH 4-10. Their high selectivity minimizes side reactions and eliminates the need for quenching steps¹. Consequently, Click-based immobilization is reliable, convenient, and operationally flexible. Due to the fast reaction kinetics, coupling under physiological conditions is feasible; however, maximum immobilization densities may be somewhat lower. Therefore, electrostatic preconcentration remains advantageous for achieving high ligand loading.

DBCO-modified surfaces exhibit a higher hydrophobicity than azide-modified counterparts or unmodified CMD coatings. This can lead to increased nonspecific binding, which is at least partially mitigated by the presence of nonionic detergents (e.g., Tween) in the running buffer. DBCO-modified sensor chips are most useful when the ligand naturally contains azide functionalities. Otherwise, the azide-modified sensor coating combined with a DBCO-modified ligand is generally preferred due to its higher intrinsic bioinertness.

Key features:

Versatile ligand coupling: Efficient and selective Click reaction enables covalent attachment of azide-functionalized ligands. Unlike classic amine coupling, ligand density can be iteratively increased after initial coupling, allowing fast optimization of immobilization levels and efficient sensor chip use.
Superior efficiency: High efficiency of the Click reaction permits ligand immobilization even under physiological conditions.
Azide functionalization: Ligands can be conveniently azide-modified using labeling strategies analogous to biotinylation; site-specific conjugation via azide-containing unnatural amino acids is also feasible.
Application versatility: Suitable for kinetic, equilibrium, and concentration analyses, as well as diverse screening applications in drug discovery.

Schematic illustration of a 3D DC sensor chip. Turquoise DBCO groups and red carboxyl groups are distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows an azide-modified antibody covalently immobilized via Click chemistry to a DBCO group.²

Product code	DCD50L	DCD200M
Base coating	3D, 30 nm bioinert CM-dextran (low density)	3D, 200 nm bioinert polycarboxylate (medium density)
Covalent immobilization capacity [µRIU]²	≈ 3,000	≈ 11,000
Recommended ligands	proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids	proteins peptides nucleic acids small molecules
Intended purpose	covalent click immobilization of azide-functionalized ligands use of detergent-containing buffers recommended	covalent click immobilization of azide-functionalized ligands (high density) use of detergent-containing buffers recommended

¹ This is valid only when the analyte lacks its own Click-reactive functionality; otherwise, unintended covalent attachment may occur.

² All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

³Covalent immobilization capacity was assessed by injecting 100 µg/mL azide-modified bovine serum albumin (BSA) in 5 mM sodium acetate buffer (pH 5.0), with 1 µRIU corresponding approximately to 1 RU.

Chloroalkane–modified sensor chips

XanTec’s HOP, HOHC, and HOD sensor chips are coated with either a bioinert 2D carboxylate (HOP) or a 3D polycarboxylate hydrogel (HOHC, HOD), each derivatized with short-chain chloroalkanes. These functional groups form stable covalent bonds with fusion proteins bearing a 33 kDa Halo tag. Reaction is rapid, highly specific, and occurs spontaneously under preconcentration conditions, ensuring uniform ligand orientation on the sensor surface.

Compared to conventional non-selective methods such as EDC/NHS activation, the Halo tag immobilization process offers substantial advantages. It is not only significantly more convenient, but also allows for a more controlled and well-defined immobilization. By avoiding random ligand orientation and ligand crosslinking, which often occur with EDC/NHS coupling, this approach results in a homogeneous ligand population, ultimately leading to improved data quality and reproducibility in kinetic and affinity assays. Consequently, experimental workflows are streamlined, requiring fewer optimization steps, while the consistency and integrity of kinetic interaction analyses are enhanced, making it easier to draw reliable conclusions about binding kinetics and affinities.

Adequate electrostatic preconcentration remains essential for successful protein immobilization. However, traditional preconcentration scouting cannot be performed because Halo-tagged fusion proteins would directly bind to the sensor chip. Therefore, preconcentration and immobilization must be evaluated simultaneously or on a separate non-derivatized sensor chip. For further details, please refer to the product manual.

Key features:

Faster assay development: No chemical activation required for covalent coupling. Ligand immobilization occurs directly under preconcentration conditions with adjustable ligand density, enabling rapid and reproducible assay setup.
Exceptional stability: Covalent coupling of Halo-tagged protein results in drift-free, permanently immobilized ligands.
Oriented immobilization: Site-directed attachment ensures maximum ligand activity.
Versatile capture capacity: Available on CMDP, CMD, and HC base coatings, offering different immobilization densities for specific experimental requirements.

Schematic illustration of a 3D HOHC sensor chip. Blue and red dots represent chloro groups and negatively charged carboxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows a Halo-tagged recombinant protein (yellow, with the HaloTag highlighted in red) covalently immobilized via a chloro group.¹

Product code	HOP	HOD200M	HOHC200M
Base coating	2D, ultra-short CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)
Immobilization capacity [µRIU]²	≈ 3,000	≈12,000	≈15,000
Recommended ligands	Halo-tagged peptides and proteins
Recommended analytes	proteins peptides nucleic acids viruses and cells	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids small molecules carbohydrates
Intended purpose	immobilization of Halo-tagged peptides and proteins use of detergent-containing buffers recommended	high-density immobilization of Halo-tagged proteins use of detergent-containing buffers recommended	high-density immobilization of Halo-tagged proteins use of detergent-containing buffers recommended

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Based on specific binding of 30 µg/mL HaloTag®-GST fusion protein (61 kDa) in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU.

HLC–modified sensor chips

XanTec’s HLC sensor chips are based on a 3D hydrogel matrix composed of highly flexible, bioinert polycarboxylate chains grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached through their amine, thiol, or aldehyde groups using established coupling chemistries such as EDC/NHS activation, thiol-maleimide coupling, or reductive amination. This versatility enables the immobilization of a wide range of biomolecules, including proteins, antibodies, peptides, nucleic acids, carbohydrates, and small organic compounds.

Compared with native HC coatings, a large fraction of carboxyl groups in the HLC polymer is converted to hydroxyl groups, substantially lowering the surface charge. As a result, HLC sensor chips are even more bioinert than their HC counterparts, and are particularly advantageous in experiments affected by persistent nonspecific binding—an issue often encountered with strongly positively charged biomolecules. The remaining carboxyl groups are typically sufficient to support electrostatic preconcentration and efficient covalent ligand coupling.

Key features:

Reduced surface charge: Significantly lower nonspecific binding than native HC, especially with highly positively charged biomolecules; particularly suited for complex matrices such as cell culture media.
Versatile ligand coupling: Covalent coupling through amine, thiol, or aldehyde groups via standard chemistries (EDC/NHS, maleimide, reductive amination).
Wide immobilization range: From several thousand to ~18,000 μRIU, suitable for analytes from whole cells and viruses to fragments < 300 Da.
Application versatility: Suitable for kinetic, equilibrium, and concentration analyses, as well as diverse screening applications in drug discovery.
High chemical stability: Withstands typical regeneration conditions, maintaining consistent response levels and kinetic behavior after multiple regeneration cycles.
No polysaccharide backbone: Lack of carbohydrate motifs prevents unwanted interactions with lectins or carbohydrate-binding proteins, making HLC sensor chips ideal for analyzing this class of biomolecules.

Schematic illustration of a charge-reduced 3D HLC sensor chip. Red and grey dots represent negatively charged carboxyl groups and charge-neutral hydroxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field.¹

Product code	HLC30M	HLC200M	HLC1500M
Base coating	3D, 30 nm bioinert polycarboxylate (medium density, reduced charge)	3D, 200 nm bioinert polycarboxylate (medium density, reduced charge)	3D, 1500 nm bioinert polycarboxylate (medium density, reduced charge)
Immobilization capacity [µRIU]²	≈ 6,000	≈ 11,000	≈ 18,000
Recommended ligands	proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids viruses and cells carbohydrates	proteins peptides nucleic acids small molecules carbohydrates	peptides nucleic acids small molecules carbohydrates
Intended purpose	kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates use with biomolecules with a high tendency for electrostatic interactions with polycarboxylates applications requiring low diffusion limitation compatible with complex buffer compositions	kinetics of small and medium analytes use with biomolecules with a high tendency for electrostatic interactions with polycarboxylates compatible with complex buffer compositions	kinetics of medium and small analytes equilibrium analysis concentration analysis use with biomolecules with a high tendency for electrostatic interactions with polycarboxylates compatible with complex buffer compositions

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

¹ Preconcentration capacity determined by injecting 100 µg/mL bovine serum albumin (BSA) in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields can vary and depend strongly on the properties of the protein to be immobilized. Under optimal conditions, typical coupling efficiencies range from approximately 20–45% of the respective electrostatic preconcentration capacity, with acidic proteins generally exhibiting lower coupling efficiencies.

HCX sensor chips (NHS–preactivated)

XanTec’s HCX sensor chips are based on a 3D hydrogel matrix composed of flexible polycarboxylate chains grafted onto a hydrophilic adhesion promoter on a gold support. The carboxylate groups are partially pre-activated with NHS esters, providing medium to high activation level and enabling high immobilization capacities. This pre-activation significantly simplifies and accelerates ligand coupling, making HCX chips a powerful tool for streamlined workflows. They are particularly advantageous for spot immobilization in imaging SPR systems.

Although a certain fraction of a spotted ligand will immobilize when drying spots, electrostatic preconcentration substantially increases immobilization yields. A preconcentration scouting step is therefore recommended prior to coupling. However, small molecules freely diffuse into the hydrogel and do not require preconcentration for efficient immobilization.

Key features:

Save time: Pre-activated sensor chips allow direct and convenient ligand immobilization, saving time and effort.
Reliable results: Avoid variability of reagent quality and instead achieve consistent, reproducible immobilization every time.
Flexibility: From moderate-capacity chips for protein-protein interaction studies to ultra-high-capacity formats for fragment screening, HCX covers the entire spectrum of ligand density requirements.

Schematic illustration of a pre-activated 3D HCX sensor chip. Yellow donut symbols and red dots represent amine-reactive NHS esters and negatively charged carboxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field.¹

Product code²	HCX30M	HCX200M	HCX1000M
Base coating	3D, 30 nm bioinert polycarboxylate (medium density) NHS pre-activated	3D, 200 nm bioinert polycarboxylate (medium density) NHS pre-activated	3D, 1000 nm bioinert polycarboxylate (medium density) NHS pre-activated
Immobilization capacity [µRIU]³	≈ 9,000	≈ 15,000	≈ 20,000
Recommended ligands⁴	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids	proteins peptides nucleic acids
Recommended analytes	proteins peptides nucleic acids viruses and cells small molecules (limited) carbohydrates	peptides small molecules nucleic acids (low-molecular-weight) carbohydrates (low-molecular-weight)	small molecules fragments carbohydrates (low-molecular-weight) nucleic acids (low-molecular-weight)
Intended purpose	spot immobilization in imaging SPR systems kinetics of medium and large analytes including carbohydrates especially suitable for weak binders with fast on- and off-rates immobilization of secondary antibodies and capture proteins	spot immobilization in imaging SPR systems kinetics of small analytes equilibrium analysis especially suitable for situations requiring higher capture densities immobilization of secondary antibodies and capture proteins	spot immobilization in imaging SPR systems equilibrium analysis fragment screening concentration analysis especially suitable for situations requiring very high immobilization densities

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² This overview represents a selection of the full HCX sensor chip portfolio.

³ Covalent immobilization capacity was assessed by injecting 100 µg/mL bovine serum albumin (BSA) in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU.

⁴ Ligands require an NHS-reactive amine.

Biotin–modified sensor chips

XanTec's biotin-modified sensor chips are coated with a bioinert (poly)carboxylate matrix (CMD or HC), pre-functionalized with a low-molecular-weight biocytin group. Owing to the exceptionally high affinity of biotin for avidin family proteins (dissociation constant ≈ 10⁻¹⁵ M for streptavidin), these chips enable highly stable immobilization with essentially negligible dissociation of bound streptavidin or streptavidin-ligand complexes.

XanTec's biotin sensor chips have gained popularity due to their compatibility with Switchavidin, an avidin derivative with a switchable affinity for biotin. This allows quantitative ligand removal from the biotin-functionalized surface, which is difficult and time-consuming when using streptavidin. In typical workflows, Switchavidin is pre-incubated with a biotinylated ligand at a controlled stoichiometry, followed by immobilization of the Switchavidin-ligand complex on the biotin sensor surface. Without this precomplex formation, Switchavidin can become buried within the hydrogel matrix, significantly reducing the effective ligand-binding capacity.

Key features:

Regenerable with Switchavidin: Enables reversible capture of biotinylated ligands via controlled binding and removal of Switchavidin.
Oriented immobilization: Precise biotinylation strategies (e.g., AviTag) support uniform ligand orientation, maximizing activity and reproducibility.
Versatile capture capacity: Available on CMD and HC base coatings, offering different capture capacities and sensor matrices to match specific experimental needs.
High chemical stability: Supports stringent regeneration conditions for the effective removal of Switchavidin, and potentially also streptavidin, from the sensor chip.

Schematic illustration of a 3D BHC sensor chip. Red and yellow/green dots represent negatively charged carboxyl groups and biotin groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows immobilized Streptavidin (purple) pre-coupled with a biotinylated ligand.¹

Product code	BP	BD200M	BHC200M
Base coating	2D, ultra-short CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)
Immobilization capacity [µRIU]²	≈ 800	≈ 7,000	≈ 7,000
Recommended ligands	biotinylated proteins peptides nucleic acids		biotinylated proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids viruses and cells	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids small molecules carbohydrates
Intended purpose	compatible with Switchavidin-mediated capture of biotinylated ligands kinetic analysis of medium and large analytes especially suitable for weak binders with fast association and dissociation rates	compatible with Switchavidin-mediated capture of biotinylated ligands kinetic analysis of medium and small analytes especially suitable for situations requiring higher capture densities	compatible with Switchavidin-mediated capture of biotinylated ligands kinetic analysis of medium and small analytes including carbohydrates especially suitable for situations requiring higher capture densities

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Based on specific binding of 100 µg/mL streptavidin in phosphate-buffered saline (PBS), with 1 µRIU corresponding approximately to 1 RU.

Sensor chips with lipophilic anchors (HPP, LP and LD)

XanTec’s LP and LD sensor chips are coated with a bioinert 2D (CMDP) or 3D (CMD500L) carboxymethyl-dextran polymer matrix, partially functionalized with lipophilic alkyl anchor groups. In contrast to solely hydrophobic 2D chip surfaces such as HPP, LP and LD sensor chips are partially hydrophilic and conserve the functionality of lipid bilayer structures.

HPP sensor chips provide a dense 2D hydrophobic coating, which allows formation of stabilized lipid monolayers. The hydrophobic surface enables lipid monolayer adsorption for studying lipid-associated interactions.

LP sensor chips consist of a hydrophilic 2D CMDP cushion functionalized with lipophilic alkyl anchors. This architecture enables reversible adsorption and rupture of lipid vesicles, resulting in the formation of a supported lipid bilayer (SLB). Membrane proteins incorporated within vesicles typically remain structurally intact and maintain biological activity after bilayer formation, making LP chips suitable for interaction studies involving moderate- to high-molecular-weight analytes, such as proteins. LP surfaces also serve as robust supports for SLB model systems, offering multiple regeneration cycles.

LD sensor chips are coated with a hydrophilic, 3D hydrogel based on brush-structured carboxymethyl-dextran. Lipid vesicles can diffuse into the LD hydrogel matrix. Incorporation of the lipophilic anchor groups allows the reversible capture of such vesicles. The shape of the vesicles usually remains intact, making them suitable for biomolecular investigation of (trans)membrane proteins such as G protein-coupled receptors (GPCRs). Compared to LP, the immobilization capacity of LD sensor chips is ≈ 50% higher, allowing investigation of biomolecular interactions, including smaller analytes.

Key features:

HPP – Hydrophobic coating: Enables formation of a supported lipid monolayer; user-prepared liposomes adsorb and reorganize spontaneously into a monolayer with hydrophilic headgroups facing the bulk solution; not suitable for transmembrane proteins (risk of denaturation upon vesicle adsorption).
LP – Supported lipid bilayer (SLB): Hydrophilic 2D coating with hydrophobic anchors supports reversible adsorption/rupture of vesicles and SLB formation; stabilization and preservation of (trans)membrane proteins for interaction analysis; regenerable surface suitable for multiple assay cycles.
LD – Vesicle immobilization in 3D hydrogel: Hydrophobic anchors enable reversible immobilization of intact lipid vesicles and associated (trans)membrane proteins (e.g., GPCRs); higher immobilization densities support investigation of small analytes; regenerable surface enables multiple assay cycles.

Schematic illustrations of XanTec’s lipophilic sensor chip variants.
HPP (left): formation of a supported lipid monolayer (hydrophilic headgroups in blue, aliphatic tails in purple) on a hydrophobic planar self-assembled alkyl monolayer (SAM, orange).
LP (center): formation of a supported lipid bilayer on a hydrophilic CMDP base coating functionalized with hydrophobic anchor groups (orange).
LD (right): immobilization and stabilization of intact lipid vesicles within a 3D CMD sensor matrix modified with hydrophobic anchor groups (orange).¹

Product code	HPP	LP	LD
Base coating	2D, hydrophobic planar alkyl layer	2D, carboxymethyl-dextran surface, partially alkyl-derivatized	3D, carboxymethyl-dextran surface, partially alkyl-derivatized
Immobilization capacity [µRIU]²	n. a.	≈ 9,000	≈ 13,000
Recommended ligands	Lipid monolayer	Supported lipid bilayer from vesicles (trans)membrane proteins	vesicles (trans)membrane proteins
Recommended analytes	proteins peptides nucleic acids viruses and cells carbohydrates	proteins peptides nucleic acids	proteins peptides nucleic acids small molecules
Intended purpose	supported lipid monolayer formation interactions of (bio)molecules with lipid monolayers and supported lipid monolayers	reversible supported lipid bilayer formation interaction analysis involving immobilization of (trans)membrane proteins in SLBs	reversible vesicle immobilization interaction analysis involving immobilization of (trans)membrane proteins applications requiring higher immobilization levels than LP sensor chips

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Capacity determined by injecting an in-house vesicle formulation, with 1 µRIU corresponding approximately to 1 RU.

Alginate–modified sensor chips

XanTec’s alginate (AL) sensor chips are based on a 50 nm alginate hydrogel matrix grafted onto a hydrophilic adhesion promoter on a gold support. The negatively charged carboxyl groups distributed along the polysaccharide chains enable efficient electrostatic preconcentration of ligands prior to covalent coupling.

Ligands can be covalently attached through their amine, thiol, or aldehyde groups using standard coupling chemistries such as EDC/NHS activation, thiol–maleimide conjugation, or reductive amination. This allows immobilization of proteins, antibodies, peptides, nucleic acids, and small molecules. Efficient protein immobilization requires electrostatic preconcentration before covalent attachment.

XanTec’s AL sensor chips were specifically developed as a functionally equivalent alternative to Bio-Rad’s ProteOn GLM sensor chips. They provide ProteOn users with a compatible alginate-based hydrogel surface and represent a reliable option when CMD- or HC-based coatings are unsuitable or cannot be applied.

In addition, AL sensor chips serve as benchmark coatings for assessing nonspecific binding during the development and evaluation of biomedical and antifouling surface chemistries.

Key features:

Alternative to ProteOn GLM sensor chips: Provides a 3D alginate-based hydrogel matrix for covalent ligand immobilization.
Versatile ligand coupling: Supports amine-, thiol-, and aldehyde-based coupling using established chemistries (EDC/NHS, maleimide, reductive amination).
Benchmark surface: Suitable as a reference coating for evaluating nonspecific binding and new biomedical coating materials.

Schematic illustration of an AL sensor chip. Red dots represent negatively charged carboxyl groups distributed along the green polysaccharide chains. The decaying red gradient represents the evanescent field.¹

Product code	AL
Base coating	3D, 50 nm alginate (medium density)
Electrostatic preconcentration capacity [µRIU]²	≈ 13,000
Recommended ligands	proteins peptides
Recommended analytes	proteins peptides nucleic acids viruses and cells
Intended purpose	alternative to ProteOn GLM sensor chips kinetic and affinity analysis of medium and large analytes benchmark surface for biomedical coating evaluation

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Preconcentration capacity determined by injecting 100 µg/mL BSA in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields depend on ligand properties and typically range from approximately 20–45% of the respective electrostatic preconcentration capacity.

C-type sensor chips

XanTec’s C-type sensor chips are based on a synthetic 3D hydrogel matrix composed of linear aliphatic polycarboxylate chains with very high charge density, grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached via their amine, thiol, or aldehyde groups using established coupling chemistries such as EDC/NHS activation, thiol–maleimide conjugation, or reductive amination. This enables immobilization of proteins, antibodies, peptides, nucleic acids, and small molecules.

The exceptionally high charge density renders the surface susceptible to nonspecific binding of cationic biomolecules; however, the aliphatic polymer backbone provides outstanding chemical robustness. This makes C-type coatings particularly well suited for demanding chemical environments and for non-biological applications that require a dense and chemically well-defined carboxylate (COOH) surface, such as solid-phase syntheses or the preparation of metal–organic frameworks (MOFs).

Key features:

Exceptional chemical stability: The aliphatic polymer backbone provides the highest chemical robustness within XanTec’s portfolio, enabling use under harsh chemical conditions and supporting non-biological applications requiring a dense COOH surface.
Very high carboxyl group density: Enables highly efficient ligand immobilization under electrostatic preconcentration conditions.
Versatile coating thickness: Available in thicknesses from 30 to 150 nm, allowing flexible adaptation to specific experimental requirements.

Schematic illustration of a 3D C-type sensor chip. Red dots represent negatively charged carboxyl groups distributed along the grey polymer chains. The decaying red gradient represents the evanescent field.¹

Product code	C30M	C80M	C150D
Base coating	3D, 30 nm aliphatic polycarboxylate (medium density)	3D, 80 nm aliphatic polycarboxylate (medium density)	3D, 150 nm aliphatic polycarboxylate (high density)
Electrostatic preconcentration capacity [µRIU]²	≈ 9,000	≈ 23,000	≈ 50,000
Recommended ligands	proteins peptides carbohydrates nucleic acids (spot immobilization)
Recommended analytes	proteins peptides nucleic acids carbohydrates	proteins peptides nucleic acids carbohydrates	peptides small molecules fragments small carbohydrates
Intended purpose	spot immobilization of ligands requiring high chemical robustness (e.g. amino-modified DNA oligonucleotides) non-biological applications requiring a dense and well-defined carboxylate (COOH) surface

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Preconcentration capacity determined by injecting 100 µg/mL bovine serum albumin (BSA) in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields depend on ligand properties and typically range from approximately 25–45% of the respective electrostatic preconcentration capacity.

CMPG–modified sensor chips

XanTec’s CMPG sensor chips are based on a short, dendritic carboxymethylated polyglycerol (CMPG) 2D sensor matrix grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached via their amine, thiol, or aldehyde groups using standard coupling chemistries such as EDC/NHS activation, thiol–maleimide conjugation, or reductive amination. This enables immobilization of proteins, antibodies, peptides, nucleic acids, and small molecules.

Although the protein immobilization capacity is relatively low (typically ≈ 1,000–2,000 µRIU), the highly hydrophilic, dendritic polymer architecture provides outstanding resistance to nonspecific binding. CMPG surfaces are therefore particularly well suited for experiments challenged by persistent nonspecific interactions, especially those involving strongly positively charged biomolecules or complex sample matrices such as cell culture media.

Key features:

Very low background binding: Excellent suppression of nonspecific binding, particularly of highly cationic biomolecules; well suited for complex sample matrices such as cell culture media.
Versatile ligand coupling: Supports covalent attachment through amine, thiol, or aldehyde groups using established chemistries (EDC/NHS, maleimide, reductive amination).
Low sensor matrix profile: The planar 2D coating enables reliable kinetic analysis of systems with fast association and dissociation rates.
No polysaccharide backbone: Absence of carbohydrate motifs prevents unwanted interactions with lectins or carbohydrate-binding proteins.

Schematic illustration of a CMPG sensor chip. Red dots represent negatively charged carboxyl groups distributed along the short green polyglycerol chains. The decaying red gradient represents the evanescent field.¹

Product code	CMPG
Base coating	2D, carboxymethylated polyglycerol (high density)
Electrostatic preconcentration capacity [µRIU]²	≈ 2,000
Recommended ligands	proteins peptides carbohydrates
Recommended analytes	proteins peptides nucleic acids viruses and cells
Intended purpose	kinetic analysis of medium and large analytes especially suitable for weak binders with fast on- and off-rates use with biomolecules engaging in strong electrostatic interactions with polycarboxylates applications requiring low diffusion limitation

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Preconcentration capacity determined by injecting 100 µg/mL BSA in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields depend strongly on ligand properties and typically range from approximately 20–45% of the respective electrostatic preconcentration capacity.

Heparin–modified sensor chips

XanTec’s heparin (HEP) sensor chips are based on a 50 nm heparin hydrogel matrix grafted onto a hydrophilic adhesion promoter on a gold support. Ligands can be covalently attached via their amine, thiol, or aldehyde groups using standard coupling chemistries such as EDC/NHS activation, thiol–maleimide conjugation, or reductive amination. Efficient protein immobilization requires electrostatic preconcentration prior to covalent coupling.

Although heparin surfaces are not typically the first choice for routine kinetic or affinity analyses, they provide a valuable alternative when CMD- or HC-based sensor chips are unsuitable. In addition, HEP sensor chips serve as benchmark coatings for assessing nonspecific binding during the development and evaluation of biomedical or antifouling surface chemistries, particularly in applications focused on hemocompatibility.

Key features:

Alternative to CMD or HC surfaces: Provides a 3D hydrogel matrix for covalent ligand immobilization when CMD or HC coatings are unsuitable.
Versatile ligand coupling: Supports covalent attachment through amine, thiol, or aldehyde groups using established chemistries (EDC/NHS, maleimide, reductive amination).
Benchmark surface: Useful as a reference coating for evaluating nonspecific binding and assessing new biomedical coating materials.

Schematic illustration of an HEP sensor chip. Red dots represent negatively charged carboxyl groups distributed along the green polysaccharide chains. The decaying red gradient represents the evanescent field.¹

Product code	HEP
Base coating	3D, 50 nm heparin (medium density)
Electrostatic preconcentration capacity [µRIU]²	≈ 11,000
Recommended ligands	proteins peptides
Recommended analytes	proteins peptides nucleic acids viruses and cells
Intended purpose	alternative to CMD50M or HC30M kinetic and affinity analyses of medium and large analytes reference surface for biomedical coating evaluation

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Preconcentration capacity determined by injecting 100 µg/mL BSA in 5 mM sodium acetate pH 5.0, with 1 µRIU corresponding approximately to 1 RU. Maximum covalent coupling yields depend strongly on ligand properties and typically range from approximately 20–45% of the respective electrostatic preconcentration capacity.

NTA–modified sensor chips (Ni)

XanTec’s NiD and NiHC sensor chips are coated with a bioinert (poly)carboxylate hydrogel matrix modified with nitrilotriacetic acid (NTA) functionalities. These sensor chips enable the reversible capture immobilization of His-tagged biomolecules through complex formation with transition metal ions, preferably Ni²⁺. For sufficient binding stability, the His-tag should contain 6–10 histidine residues. Complete regeneration of both NiD and NiHC surfaces is typically achieved by removal of chelated Ni²⁺ ions using chelating agents (e.g., EDTA) or competitive ligands such as imidazole.

Binding characteristics:
NiD sensor chips exhibit predominantly monovalent interactions between NTA(Ni²⁺) complexes and His-tags, resulting in dissociation rates (k_off) in the range of 10⁻³ s⁻¹.
NiHC sensor chips support multivalent binding due to the flexibility of the polymer chains, increasing binding stability by up to three orders of magnitude. Baselines after His-tagged ligand capture show minimal to no drift, with typical k_off values of 10⁻⁵–10⁻⁶ s⁻¹.¹

A known limitation of NTA-based capture surfaces is the potential for nonspecific interactions between chelated Ni²⁺ ions and protein analytes or complex sample matrices (e.g., FBS). Proteins containing exposed histidines or metal-affinity motifs may bind non-selectively, increasing background signals. For assays sensitive to nonspecific binding, alternative immobilization strategies (e.g., anti-His-tag antibody capture) should be considered.

Key features:

Fast assay development: No preconcentration or surface activation required; reversible ligand immobilization under physiological conditions.
Easy regeneration: Chip surfaces regenerated using EDTA or imidazole injections.
Outstanding stability (NiHC): Multivalent binding minimizes baseline drift during analysis.
Oriented immobilization: His-tag–directed capture ensures maximum ligand activity.
Versatile capture capacity: Available on CMD and HC base coatings for a broad range of analytes.

Schematic illustration of a 3D Ni sensor chip. Green and red dots represent NTA-bound Ni²⁺ ions and negatively charged carboxyl groups distributed along the green polymer chains. The decaying red gradient represents the evanescent field. The magnified view shows an NTA-bound Ni²⁺ ion coordinating a protein ligand (yellow) via its His-tag (light blue).²

Product code	NiP	NiD200M	NiHC200M	NiHC1500M
Base coating	2D, ultra-short bioinert CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)	3D, 1500 nm bioinert polycarboxylate (medium density)
Capture immobilization capacity [µRIU]⁴	≈ 120	≈ 400	≈ 1,200	≈ 2,000
Ligands	His-tagged peptides and proteins
Recommended analytes	proteins large peptides large nucleic acids	proteins peptides nucleic acids	nucleic acids small molecules peptides carbohydrates	nucleic acids small molecules small carbohydrates
Intended purpose	suitable for weak binders recommended in combination with EDC/NHS stabilization	equivalent to competitor NTA chips suitable for weak binders recommended in combination with EDC/NHS stabilization	ideal for medium to small analytes higher capture densities minimal baseline drift	maximum capture capacity optimized for small analytes minimal baseline drift

¹ Maximum binding stability requires immobilization at approximately one-third of the chip’s maximum capture capacity.

² All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

³ Table includes a selection from XanTec’s full Ni sensor chip portfolio.

⁴ Binding capacities are based on the capture level of His₆-peptide from a 5 µM solution after a 180 s stabilization period, with 1 µRIU corresponding approximately to 1 RU. Immobilization capacities of His-tagged proteins are considerably higher.

Protein A–modified sensor chips

XanTec’s Protein A (PA) sensor chips are coated with a bioinert polycarboxylate matrix pre-functionalized with a recombinant 47 kDa Protein A. This non-glycosylated variant contains five IgG-binding domains, providing high-affinity binding to the Fc region of all human IgG1, IgG2, and IgG4 subclasses. It also shows strong affinity for mouse IgG2a, IgG2b, and IgG3, as well as total IgG from cow, goat, sheep, rabbit, and other mammals.

Due to the slightly hydrophobic character of Protein A, some nonspecific binding may occur. The use of blocking agents (e.g., BSA) and/or nonionic detergents (e.g., Tween) in the running buffer is therefore recommended to minimize background signals. XanTec’s PA sensor chips are ready-to-use, eliminating time-intensive assay optimization and streamlining workflows.

Key features:

Fast assay development: No preconcentration or surface activation required; reversible capture under physiological conditions.
Simple regeneration: Regeneration using glycine·HCl pH 1.5–2.5.
High stability: Robust capture surfaces enabling multiple reuse cycles.
Oriented immobilization: Directed Fc-region binding ensures high ligand activity.

Schematic illustration of a 3D PA sensor chip. Red dots represent negatively charged carboxyl groups along the green polymer chains. The magnified view shows Protein A (orange) binding to the Fc region of an antibody (blue).¹

Product code	PAP	PAD200L	PAHC30M	PAHC200M
Base coating	2D, ultra-short bioinert CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (low density)	3D, 30 nm bioinert polycarboxylate (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)
Capture immobilization capacity [µRIU]³	≈ 5,000	≈ 12,000	≈ 6,000	≈ 11,000
Ligands	Fc region of human IgG₁, IgG₂, and IgG₄; strong affinity for mouse IgG_2a, IgG_2b, IgG₃, and total IgG from cow, goat, sheep, and rabbit
Recommended analytes	proteins peptides nucleic acids	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids carbohydrates	proteins peptides nucleic acids carbohydrates small molecules
Intended purpose	kinetic measurements via oriented Fc capture	capture of Fc fusion proteins and antibodies for medium- to large-sized analytes antibody quantification	analysis of small- to medium-sized analytes antibody quantification	analysis of medium- to large-sized analytes including carbohydrates

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Table includes a selection from XanTec’s full PA sensor chip portfolio.

³ Maximum immobilization capacities are based on capture from a 100 µg/mL solution of Protein A-purified, pooled rabbit IgG in PBS, with 1 µRIU corresponding approximately to 1 RU.

Protein AG–modified sensor chips

XanTec’s Protein AG (PAG) sensor chips are coated with a bioinert polycarboxylate matrix pre-functionalized with a recombinant 50.5 kDa Protein AG. This non-glycosylated variant combines four IgG-binding domains from Protein A and two IgG-binding domains from Protein G, providing high-affinity binding to the Fc region of all human IgG subclasses. In addition, it exhibits strong affinity for mouse IgG2a, IgG2b, and IgG3, as well as total IgG from cow, goat, sheep, rabbit, and other mammals.

To ensure highly specific IgG binding, non-essential domains (e.g., cell wall-, cell membrane-, and albumin-binding regions) have been removed from the recombinant Protein AG. Nevertheless, due to its slightly hydrophobic character, some nonspecific binding may occur. The inclusion of blocking agents (e.g., BSA) and/or nonionic detergents (e.g., Tween) in the running buffer is therefore recommended to minimize background signals. XanTec’s PAG sensor chips are ready-to-use, enabling rapid assay setup without time-intensive surface optimization.

Key features:

Fast assay development: No preconcentration or surface activation required; controlled and reversible capture under physiological conditions.
Broad antibody compatibility: Wider subclass and species coverage than Protein A.
Simple regeneration: Regeneration using glycine·HCl pH 1.5.
High stability: Robust capture surfaces support repeated regeneration cycles.
Oriented immobilization: Directed Fc-region binding ensures high ligand activity.

Schematic illustration of a 3D PAG sensor chip. Red dots represent negatively charged carboxyl groups distributed along the green polymer chains. The magnified view shows Protein AG (yellow–orange) binding to the Fc region of an antibody (blue).¹

Product code	PAGP	PAGD200L	PAGHC30M	PAGHC200M
Base coating	2D, ultra-short bioinert CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (low density)	3D, 30 nm bioinert polycarboxylate (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)
Capture immobilization capacity [µRIU]³	≈ 5,000	≈ 12,000	≈ 6,000	≈ 11,000
Ligands	Fc region of all human IgG subclasses; strong affinity for mouse IgG_2a, IgG_2b, IgG₃, and total IgG from cow, goat, sheep, and rabbit
Recommended analytes	proteins peptides nucleic acids	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids carbohydrates	proteins peptides nucleic acids carbohydrates small molecules
Intended purpose	kinetic measurements based on oriented Fc-region immobilization	capture immobilization of Fc fusion proteins and antibodies for medium- to large-sized analytes antibody quantification	capture immobilization of Fc fusion proteins and antibodies for small- to medium-sized analytes antibody quantification	capture immobilization of Fc fusion proteins and antibodies for medium- to large-sized analytes including carbohydrates antibody quantification

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Table includes a selection from XanTec’s full PAG sensor chip portfolio.

³ Maximum immobilization capacities are based on capture from a 100 µg/mL solution of Protein A-purified, pooled rabbit IgG in PBS, with 1 µRIU corresponding approximately to 1 RU.

RG sensor chips (regenerable DNA-mediated capture)

XanTec’s RGD200M sensor chips are coated with a bioinert carboxymethyl-dextran matrix derivatized with a 24-nt single-stranded DNA oligonucleotide. This surface enables DNA-mediated, reversible ligand immobilization via hybridization with a complementary DNA sequence. Captured ligands are stably bound under physiological conditions, while quantitative regeneration is achieved by alkaline denaturation of the double-stranded DNA, restoring the surface for subsequent capture cycles.

Two complementary immobilization strategies are available. In Variant 1 (RG-SA), streptavidin pre-conjugated with the complementary DNA oligo is first immobilized via DNA hybridization, followed by capture of biotinylated ligands. In Variant 2 (RG-Modifier), ligands are directly conjugated to the complementary DNA oligo (e.g., via DBCO Click chemistry) and immobilized without an intermediate streptavidin step. Variant 2 can reduce nonspecific binding, lower diffusion limitations, enable higher capture densities, and facilitate faster assay setup.

Key features:

Very fast assay development: No preconcentration, no surface activation, and no regeneration screening required; immobilization under physiological conditions.
Maximum flexibility: Regenerable alternative to classic streptavidin/biotin capture immobilization.
Exceptional stability: Drift-free immobilization of oligo-modified streptavidin.
Simple regeneration: Quantitative regeneration enabling >100 capture/regeneration cycles.
Oriented immobilization: Controlled biotinylation (e.g., AviTag™) supports high ligand activity and reproducibility.
Ideal for large screening campaigns: Consistent surface performance across many cycles and sensor chips.

Schematic illustration of an RGD200M sensor chip. Variant 1 (left): DNA–streptavidin conjugate (purple) immobilized via hybridization captures biotinylated ligands (yellow). Variant 2 (right): DNA–ligand conjugates are directly immobilized via DNA hybridization. The decaying red gradient represents the evanescent field.¹

Product code	RGD200M	RGD200M
Base coating	3D, 200 nm bioinert CM-dextran (medium density)
Variant	Variant 1: RG-SA	Variant 2: RG-Modifier
Capture immobilization capacity [µRIU]	≈ 3,500²	≈ 5,000³
Recommended ligands	biotinylated proteins or peptides	proteins or peptides with NHS-reactive amino groups
Recommended analytes	proteins peptides	proteins peptides small molecules
Intended purpose	kinetic and equilibrium analyses of medium analytes assay development with new or uncharacterized binding partners screening of biotinylated peptide and protein libraries time-critical SPR experiments systems that are difficult to regenerate not recommended for DNA-binding or intercalating molecules	kinetic and equilibrium analyses of small, medium, and large analytes higher ligand densities than RG-SA superior baseline stability applications with difficult regeneration not recommended for DNA-binding or intercalating molecules

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Based on capture of 100 µg/mL biotinylated BSA in PBS (RG-SA pre-capture), with 1 µRIU corresponding approximately to 1 RU.

³ Based on capture of 200 nM RG-SA on an RGD200M sensor chip.

Streptavidin–modified sensor chips

XanTec’s streptavidin-modified (SA) sensor chips are coated with a bioinert (poly)carboxylate matrix pre-functionalized with a recombinant 52 kDa streptavidin tetramer. The immobilized streptavidin captures biotinylated biomolecules—including proteins, peptides, nucleic acids, and other biotin-tagged ligands—efficiently under physiological conditions. Owing to the exceptionally high affinity of streptavidin for biotin (dissociation constant ≈ 10⁻¹⁵ M), SA sensor chips enable highly stable ligand immobilization with negligible dissociation.

A general limitation of streptavidin-based capture surfaces is that the maximum achievable ligand density is typically lower than that obtained by direct covalent immobilization strategies combined with electrostatic preconcentration. This is because the pre-immobilized streptavidin tetramer occupies a significant fraction of the accessible volume within the evanescent field. This should be considered when high ligand densities are required, for example in interaction analyses involving small analytes.

Key features:

Fast assay development: No preconcentration or surface activation required; controlled ligand immobilization under physiological conditions.
Exceptional stability: The strong streptavidin–biotin interaction renders the surface resistant to most common regeneration protocols.
Oriented immobilization: Controlled biotinylation strategies (e.g., AviTag™) ensure high ligand activity and reproducibility.
Broad application range: Suitable for protein interaction studies, nucleic acid hybridization, antibody screening, and other biosensing applications.
Versatile capture capacity: Available on various CMD and HC base coatings to match specific experimental needs.

Schematic illustration of a 3D SA sensor chip. Red dots represent negatively charged carboxyl groups distributed along the green polymer chains. The magnified view shows immobilized streptavidin tetramers (purple) capturing a biotinylated protein ligand (yellow).¹

Product code	SAP	SAD200M	SAD700L	SAHC30M	SAHC200M	SAHC1500M
Base coating	2D, ultra-short CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 700 nm bioinert CM-dextran (low density)	3D, 30 nm bioinert polycarboxylate (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)	3D, 1500 nm bioinert polycarboxylate (medium density)
Capture immobilization capacity [µRIU]³	≈ 600–1,200	≈ 4,000–5,000	≈ 6,000–8,000	≈ 1,200–1,800	≈ 3,500–5,000	≈ 4,500–6,000
Recommended ligands	biotinylated proteins peptides nucleic acids			biotinylated proteins peptides nucleic acids carbohydrates
Recommended analytes	proteins peptides nucleic acids viruses and cells	proteins peptides nucleic acids small molecules (limited)	peptides nucleic acids small molecules	proteins peptides nucleic acids carbohydrates	proteins peptides nucleic acids small molecules (limited) carbohydrates	peptides nucleic acids small molecules small carbohydrates
Intended purpose	kinetics of medium and large analytes especially suitable for weak binders capture of biotinylated secondary antibodies	all-purpose surface kinetics of medium and small analytes alternative to Cytiva SA sensor chips	kinetics of small analytes equilibrium analysis very high capture densities	kinetics of medium and large analytes including carbohydrates capture of biotinylated secondary antibodies	all-purpose surface kinetics of medium and small analytes including carbohydrates high capture densities	kinetics of small analytes including carbohydrates equilibrium analysis very high capture densities

¹ All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

² Table includes a selection from XanTec’s full SA sensor chip portfolio.

³ Based on capture of 100 µg/mL biotinylated BSA in PBS, with 1 µRIU corresponding approximately to 1 RU.

SpyCatcher–modified sensor chips

XanTec’s SpyCatcher (SPY) sensor chips are coated with a bioinert (poly)carboxylate matrix pre-functionalized with recombinant 12.8 kDa SpyCatcher3™. Immobilized SpyCatcher3 forms a highly stable covalent isopeptide bond with Spy-tagged proteins under physiological conditions (pH 5–8, 4–37 °C), enabling site-directed and permanent ligand immobilization.

Compared with the widely used streptavidin/biotin system, the SpyCatcher/SpyTag technology offers two major advantages. First, the interaction is truly covalent, resulting in exceptional long-term stability and drift-free baselines. Second, SpyCatcher3 is considerably smaller than streptavidin, occupying less volume within the hydrogel matrix and allowing higher ligand capture densities. These properties make SPY sensor chips particularly attractive for applications involving short peptides, small molecules, and fragments.

SPYD200M and SPYHC200M sensor chips provide high immobilization capacities for dense ligand presentation, while SPYP sensor chips combine lower capacity with excellent diffusion characteristics typical of 2D coatings. This makes SPYP surfaces especially suitable for protein–protein interaction studies and systems with fast association and dissociation rates.

Key features:

Fast assay development: No preconcentration or chemical activation required; ligand immobilization occurs directly under physiological conditions.
Exceptional stability: Covalent SpyCatcher/SpyTag coupling ensures permanently immobilized, drift-free ligands.
Oriented immobilization: Site-directed attachment via N- or C-terminal SpyTag maximizes ligand activity.
Versatile capture capacity: Available on CMDP, CMD, and HC base coatings to match specific experimental requirements.

Schematic illustration of a 3D SPY sensor chip. Red dots represent negatively charged carboxyl groups along the green polymer chains. The magnified view shows immobilized SpyCatcher (red) covalently binding a Spy-tagged protein ligand (SpyTag in blue, protein ligand in yellow).²

Product code	SPYP	SPYD200M	SPYHC200M
Base coating	2D, ultra-short CM-dextran (high density)	3D, 200 nm bioinert CM-dextran (medium density)	3D, 200 nm bioinert polycarboxylate (medium density)
Capture immobilization capacity [µRIU]³	≈ 100	≈ 500	≈ 500
Recommended ligands	Spy-tagged peptides and proteins
Recommended analytes	proteins peptides nucleic acids viruses and cells	proteins peptides nucleic acids small molecules	proteins peptides nucleic acids small molecules carbohydrates
Intended purpose	immobilization of Spy-tagged proteins kinetics of medium and large analytes especially suitable for weak binders with fast on- and off-rates	immobilization of Spy-tagged proteins kinetics of medium and small analytes equilibrium and concentration analysis high capture densities	immobilization of Spy-tagged proteins kinetics of medium and small analytes including carbohydrates equilibrium and concentration analysis high capture densities

¹ SpyCatcher3 carries a His-tag that may interact with certain ligands or analytes.

² All illustrations are schematic representations and are not drawn to scale; dimensions, densities, and spatial relationships do not reflect actual physical or chemical proportions.

³ Based on capture of 100 nM SpyTag, with 1 µRIU corresponding approximately to 1 RU.

Agarose

Agarose (MW approx. 120 kDa) monolayer grafted to a thin adhesion promoter.

Linear polymer consisting of alternating D-galactose and 3,6-anhydro-L-galactopyranose linked by α-(1→3) and β-(1→4) glycosidic bonds.

Alginate

Alginate (or alginic acid) is a biopolymer that has found numerous applications in biomedical science and engineering. Alginate hydrogels are particularly attractive in wound healing, drug delivery, and tissue engineering applications, as these gels retain structural similarity to the extracellular matrices in tissues.

Alginate is a linear polymer composed of (1-4)-linked β-D-mannuronate (M) and its C-5 epimer α-L-guluronate (G), covalently linked together in different sequences. The monomers can appear in homopolymeric blocks of consecutive G-residues (G-blocks), consecutive M-residues (M-blocks), or alternating M and G-residues (MG-blocks). Alginates are refined from brown seaweeds of the class Phaeophyceae. As a natural product, alginates from different species of seaweed have variations in their chemical structure, resulting in different physical properties.

Click chemistry (AZC, AZD, DCD)

Click Chemistry or Click Coupling has gained increasing popularity among chemists because of its outstanding reaction kinetics, excellent selectivity, and superior stability in physiological environments. To make this bioconjugation technology available for ligand immobilization in SPR biosensing, polycarboxylate sensor coatings are modified with a very small, bioinert, azido (N3) group, while the ligand is labeled with a low-molecular-weight cyclooctyne linker (DBCO), a procedure comparable to ligand biotinylation. These groups can react in a quantitative and highly-selective manner via a simple alkyne–azide cycloaddition, forming a stable, covalent triazole linkage.

Some expression systems are capable of site-specific integration of non-natural amino acids containing an azide moiety. For efficient, selective and site-directed Click Coupling of such ligands XanTec also offers DBCO-modified sensor chips.

You can find more information about azide- and DBCO-modified polycarboxylate sensor chips for Click Chemistry here.

Linear polycarboxylate (C)

The robust C hydrogel consists of a hydrophilic, strictly linear polycarboxylate with a polyalkyl backbone. Compared with CM-Dextran (CMD) coatings, this hydrogel shows significantly lower diffusion limitation, resulting from its small molecular footprint and higher flexibility of the polymer chains. It has a high carboxyl group density and is a good choice if the ligand and analyte are small and high immobilization density is required. Due to the high negative charge density, the C coating is particularly well suited for acidic analytes, such as DNA. Harsh conditions are tolerated as the C–C backbone withstands practically all aqueous buffers and organic solvents except strong oxidizing agents.

Chondroitin sulfate (CDS)

CDS is an unbranched glycosaminoglycan composed of a chain of alternating sugars (N-acetyl-D-galactosamine and D-glucuronic acid) where each sugar can be sulfated in variable positions and quantities.

As a mucopolysaccharide and component of cartilage tissue, CDS shows good biocompatibility while being mechanically relatively rigid. The abundant carboxyl groups allow for activation using standard EDC/NHS chemistry.

CM-Cellulose

Carboxymethyl cellulose (CM-Cellulose, CMC) is a cellulose derivative with carboxymethyl groups (–CH2–COOH) bound to hydroxyl groups of the glucopyranose monomers that make up the cellulose backbone. Dependent on the degree of carboxymethylation and the polymer length, functional properties of CMC vary. CMC sensor surfaces can be useful for interaction studies of plant derived biomolecules or cellulose-related research.

Carboxymethyldextran (CMD)

Dextran and its derivative CM-Dextran have a long history as biocompatible materials and the crosslinked form of CMD (Sephadex®) has been used for more than 50 years as a stationary phase for chromatography of biomolecules. These polysaccharides are relatively bioinert and moderate flexibility of the polymer chains gives immobilized ligands a degree of freedom within the hydrogel. Dextran-based hydrogel coatings thus provide a good environment for many biomolecular interactions. The polymer main chain consists of glucose monomers with α-1,6 glycosidic linkages and occasional branches formed by α-1,3 forks. Carboxymethylation—usually one COOH per anhydroglucose unit—introduces negatively charged carboxyl groups into the dextran, which can be activated with EDC/NHS and allow for a range of secondary derivatizations, such as amino, thiol or azide groups.

CM-Dextran coatings also serve as the basis for further sensor chip types or modifications, such as Protein A/G for capture of antibodies, NTA for His₆-tags, streptavidin for biotinylated proteins, and lipophilic anchors for immobilization of vesicles and micelles.

Hydrogels from CM-Dextran coatings are suitable for many applications. However, in interaction measurements with other carbohydrates or with highly glycosylated proteins, nonspecific interactions can be high, making CMD less suitable for such analytes/ligands. Furthermore, carbohydrate-based polymers are quite bulky, occupy a significant fraction of the evanescent field, and can cause diffusion artifacts.

Planar carboxymethyldextran (CMDP)

Short carboxymethyldextran (CMD) chains in train conformation, i.e. attached almost flat to a thin adhesion promoter layer. The dextran matrix of this <5 nm thin composite coating has the same molar carboxymethylation level as our 3-dimensional CMD sensor chips, i.e. one carboxymethyl group carried per anhydroglucose unit.

The hydrated yet thin structure of this coating effectively suppresses nonspecific interactions while avoiding steric effects, which is advantageous when working with large molecules (MW >200 kDa). The CMDP coating is also suitable for particulate analytes, such as viruses and whole cells. As for all carboxyl functionalized coatings, this surface is compatible with standard EDC/NHS immobilization chemistry.

Carboxymethylpolyethyleneglycol (CMPEG)

Carboxyl-terminated polyethylene glycol (4 kDa) terminally grafted to a hydrophilic adhesion promoter. Low non-specific binding and limited negative charge density (only one COO- functionality per PEG molecule). Immobilization of biomolecules via, e.g., EDC/NHS chemistry familiar from more common chip coatings like carboxymethyldextran (CMD).

Low-molecular-weight PEG coatings combine good entropic stabilization with the advantage that the interactions take place relatively close to the gold surface, i.e., in the most sensitive part of the evanescent field. This surface is an alternative for interaction analysis of medium- or high-MW analytes if dextran- or polycarboxylate-based surfaces cannot be used.

Sulfonated polycarboxylate hydrogel (CS)

The partially sulfonated “C“-matrix consists of a biocompatible, hydrophilic, strictly linear polycarboxylate comprising of a brush-structured polyalkyl polymer backbone in which some carboxylic groups have been replaced by more acidic sulfonates. Having a lower pKa compared with carboxylates, these sulfonate groups allow preconcentration of proteins with pI <4.25. The typical characteristics of the C matrix, such as reduced diffusion limitation for bigger molecules and higher flexibility of the polymer chains compared to polysaccharide-based hydrogels, are further advantages of these coatings.

Dextran (D)

Coatings of brush-structured, unmodified dextran (D) are relatively bioinert and electrostatically neutral. The flexibility of the unbranched polymer chains gives immobilized ligands a degree of freedom. Dextran-based hydrogels thus provide an excellent environment for many biomolecular interactions. The polymer main chain consists of α-1,6 glycosidic linkages between glucose monomers, with branches from α-1,3 linkages. In contrast to the usually employed carboxymethyl dextran (CMD), the dextran coating has no charge. Due to the lack of carboxylic groups, D coatings are not compatible with standard EDC/NHS immobilization chemistry. Alternative covalent immobilization methods include oxidation to polyaldehydes and subsequent Schiff base coupling/reductive amination or (bis)epoxy activation. As there will not be any electrostatic preconcentration, high ligand concentrations are necessary for sufficient immobilization.

Gelatin

Gelatin is an irreversibly hydrolyzed form of collagen with a broad molecular weight distribution of the polymer strands. Gelatin contains 19 amino acids, predominantly glycine, proline and hydroxyproline (>50%).

HC linear polycarboxylate

To minimize steric hindrance, diffusion artifacts and nonspecific binding to carbohydrate motifs, XanTec has made efforts to substitute polysaccharides such as CM-Dextran with a better defined polycarboxylate with excellent bioinertness and smaller molecular footprint compared with the relatively bulky saccharides. The linear, unbranched, highly hydrated polymer chains of the HC hydrogel are extremely flexible; thus, they give immobilized ligands a high degree of freedom and the ability to interact in a cooperative manner with larger analytes.

In addition to these biophysical advantages, the HC coatings contain no hydroxyl groups. Therefore, they cannot form ester crosslinks upon EDC/NHS activation, a common side reaction with carboxymethylated carbohydrates. The HC polymers are fully compatible with all immobilization chemistries available for CM-Dextran sensor chips.

Structural differences between surface-grafted hydrogels based on polysaccharide (left) and HC polycarboxylate (right).

Heparin (HEP)

Heparin is a carbohydrate of the glycosaminoglycan family and consists of variably sulfated repeating disaccharide units. The most common disaccharide unit is composed of a 2-O-sulfated iduronic acid and 6-O-sulfated, N-sulfated glucosamine. Heparin as a sensor chip coating exhibits good bioinertness and can be useful in hematology research as it binds selectively plasma coagulation proteins, nucleic acid enzymes, lipases, and other proteins with specific heparin binding sites like lipoproteins and matrisome-associated proteins (e.g. growth factors and cytokines).

Hydrophobic planar alkyl layer

Dodecanethiol self-assembled monolayer. Contact angle >100°.

This flat, hydrophobic sensor surface facilitates adsorption of lipid monolayers for analysis of interactions involving lipid components. Liposomes adsorb directly onto the sensor surface and form a lipid monolayer with “tails” directed toward the hydrophobic alkyl layer and hydrophilic “heads” directed toward the solution. Unlike the usual strategy for immobilization of biomolecules, this surface has no functional groups suitable for forming covalent bonds.

Hyaluronic acid (HY)

yaluronic acid is a strongly hydrated, anionic, nonsulfated glycosaminoglycan, which makes it unique among glycosaminoglycans as they are usually sulfated. The molecular weight of the hyaluronic acid used for the HY coatings lies between 2 and 4 MDa, which corresponds to 15,000–25,000 disaccharide monomers. The hyaluronic acid used is of animal origin (chicken).

Hyaluronic acid is a polymer of disaccharides, themselves composed of D-glucuronic acid and N-acetyl-D-glucosamine, linked via alternating β-(1→4) and β-(1→3) glycosidic bonds. Due to its high biocompatibility and common presence in the extracellular matrix of tissues, hyaluronic acid has been used as the basis for biomaterial scaffolds in tissue engineering.

As hyaluronic acid (HY) hydrogels are bioinert they can be a good choice for use in experiments with ligands/analytes that are prone to nonspecific interactions with CMD and HC surfaces. Further, they can be useful for biocompatibility research and tissue engineering.

Due to the abundant carboxyl groups, HY coatings are fully compatible with all immobilization chemistries available for CM-Dextran sensor chips.

Hydrazide modified linear polycarboxylate (HZHC)

Based on the linear polycarboxylate HC with its superior biocompatibility and diffusion properties, the hydrazide modification of this surface coating offers mild and covalent immobilization of oxidized (poly)saccharides, antibodies through their oxidized carbohydrate side chains, or other molecules with aldehyde or ketone functional groups. The resulting hydrazone bond is stable if formed from ketones; if an aldehyde is used, it is recommended to stabilize the double bond by reduction.

Lipophilic anchors (LD)

The 3D hydrogel LD surface—based on 500-nm carboxymethyl dextran coating—has been optimized to capture whole vesicles and liposomes. Covalently attached to a hydrophilic polymer, lipophilic alkyl chains act as “anchors” for particulates containing lipid membranes.

Planar lipophilic anchors (LP)

On the 2D LP surface, lipophilic alkyl chains are linked to a planar carboxymethyl dextran layer (CMDP surface). In contrast to the hydrophobic alkyl SAM-modified surface (HPP) that immobilizes lipid monolayers, the LP chip is the ideal substrate to prepare supported lipid bilayers. Such 2D structures can be simply prepared by vesicle spreading and mimic the cell membrane. LD and LP surfaces are suitable for analysis of membrane protein–protein interactions or membrane protein–inhibitor interactions.

NTA immobilized on CMD (NiD)

Nitrilotriacetic acid (NTA)-derivatized carboxymethyldextran allows immobilization of polyhistidine (His₆)-tagged molecules via Ni²⁺/NTA chelation. Obvious advantages of this immobilization method are the site-directed, homogenous immobilization of ligands; capture in physiological conditions without significant changes of ionic strength and/or pH; and the ability to regenerate the sensor chip using chelators such as EDTA (350 mM) or imidazole (50 mM) followed by Ni²⁺ re-loading and immobilization of fresh ligand.

Although this approach is quite popular, a common problem with NTA sensor chips based on carboxymethyldextran is constant leaching of bound ligand from the chip surface, which results in a drifting baseline. If surfaces with a stable baseline are required, we recommend the use of our high-affinity poly-NTA sensor chips.

The use of 50 µM EDTA in running and sample buffers is recommended, as it decreases negative effects caused by contaminating heavy metal ions. Low concentrations of surfactant help to decrease nonspecific binding. HEPES buffer gives better results than Tris or PBS.

Multidentate poly-nitrilotriacetic acid (NTA)-derivatized linear polycarboxylate (NiHC)

NTA-derivatized carboxymethyldextran (CMD) allows immobilization of polyhistidine (His₆)-tagged molecules via Ni²⁺/NTA chelation. Obvious advantages of this immobilization method are the site-directed, homogenous immobilization of ligands; capture in physiological conditions without significant changes of ionic strength and pH; and the ability to regenerate the sensor chip using chelators such as EDTA (350 mM) or imidazole (50 mM) followed by Ni²⁺ re-loading and immobilization of fresh ligand.

A common problem of NTA-derivatized CMD surfaces is the constant leaching of captured ligand from the chip surface, which results in a drifting baseline. Such drift effects can easily exceed the specific signal and thus represent a major problem, especially when screening small molecules. To address this unwanted effect, XanTec has developed a poly-NTA sensor chip hydrogel coating (HC) based on a flexible polycarboxylate polymer backbone. Compared with the standard CMD–NTA chemistry, these hydrogels, which are available in 30-, 200-, 1000-, and 1500-nm thickness, improve the immobilization stability of captured His₆-tagged ligands by 2–3 orders of magnitude (Fig. 1). The higher stability of the bond results from a cooperative effect of several, closely spaced NTA groups.

Therefore, when selecting a sensor chip for immobilization of His₆-tagged proteins in critical applications with small molecule analytes, the HC variant should always be preferred to the CMD-based chip. However, the increased density of NTA groups in the HC variant may lead to higher non-specific binding (NSB). Thus, if the analyte is a large protein, or the sample matrix is complex (e.g., it contains serum proteins), the choice should be a CMD-based NTA chip to avoid NSB.

Figure 1. Interaction/affinity map of His₆-tagged A/G fusion protein with mono-NTA-derivatized carboxymethyl dextran (top) and XanTec’s poly-NTA sensor chip (bottom; NiHC1000M). Affinity increases from the upper-right corner to the lower-left corner. The affinity map was calculated from association/dissociation data using EvilFit⁵.

Nieba, L., Nieba-Axmann, S. E., Persson, A., Hämäläinen, M., Edebratt, F., Hansson, A., ... & Plückthun, A. (1997). BIACORE analysis of histidine-tagged proteins using a chelating NTA sensor chip. Analytical biochemistry, 252(2), 217-228.
Oshannessy, D. J., Odonnell, K. C., Martin, J., & Brighamburke, M. (1995). Detection and quantitation of hexa-histidine-tagged recombinant proteins on western blots and by a surface plasmon resonance biosensor technique. Analytical biochemistry, 229(1), 119-124.
Gorshkova, I. I., Svitel, J., Razjouyan, F., & Schuck, P. (2008). Bayesian analysis of heterogeneity in the distribution of binding properties of immobilized surface sites. Langmuir, 24(20), 11577-11586.

Protein A/G on carboxymethyldextran sensor chips

Recombinant fusion protein A/G (Mw around 50 kDa) is pre-immobilized either on a planar surface or in a 3-dimensional hydrogel consisting of carboxymethyldextranCMD). Recombinant protein A/G on these sensor chips consists of seven IgG-binding domains (EDABC–C2C3); the protein A portion (segments E, D, A, B, and C) is from Staphylococcus aureus, and the protein G portion (segments C2 and C3) is a Streptococcal cell wall protein. Sensor surfaces with protein A/G are designed to bind antibodies (IgGs) from different subclasses and species through the heavy chain in the Fc region. The two proteins differ in their binding properties for antibodies from different species and IgG subclasses: protein A is generally preferred for rabbit, pig, dog, and cat IgG, while protein G has better binding capacity for a broader range of mouse and human IgG subclasses (IgG1, IgG2, etc.).

The oriented and reversible immobilization of antibodies ensures high activity of the immobilized IgG without lengthy optimization of immobilization protocols or regeneration conditions. With a K_a of approximately 10⁸ M⁻¹ protein A/G–IgG complexes are sufficiently stable to yield surfaces with low drift that are suitable for a broad range of applications. Binding to fusion protein A/G is less pH-dependent than to either protein A or protein G alone. However, incubation with denaturing buffers or at slightly acidic or alkaline pH will break the interaction and results in quantitative regeneration of the chip surface, which can be repeated several hundred times.¹

J. Quinn, P. Patel, B. Fitzpatrick, B. Manning, P. Dillon, S. Daly, R. O’Kennedy, M. Alcocer, H. Lee, M. Morgan and K. Lang, Biosens. Bioelectron., 1999, 14, 587–595.

Protein A/G on linear polycarboxylate polymer (HC)

Recombinant fusion protein A/G (Mw around 50 kDa) is pre-immobilized in a 3-dimensional hydrogel consisting of a linear polycarboxylate with higher flexibility in its polymer chains than carboxymethyldextran. Recombinant protein A/G on these sensor chips consists of seven IgG-binding domains (EDABC–C2C3); the protein A portion (segments E, D, A, B, and C) is from Staphylococcus aureus, and the protein G portion (segments C2 and C3) is is a Streptococcal cell wall protein. Sensor surfaces with protein A/G are designed to bind antibodies (IgGs) from different subclasses and species through the heavy chain in the Fc region. The two proteins differ in their binding properties for antibodies from different species and IgG subclasses: protein A is generally preferred for rabbit, pig, dog, and cat IgG, while protein G has better binding capacity for a broader range of mouse and human IgG subclasses (IgG1, IgG2, etc.).

J. Quinn, P. Patel, B. Fitzpatrick, B. Manning, P. Dillon, S. Daly, R. O’Kennedy, M. Alcocer, H. Lee, M. Morgan and K. Lang, Biosens. Bioelectron., 1999, 14, 587–595.

Pectin (PC)

Pectins, also known as pectic polysaccharides, are structural heteropolysaccharides contained in the primary cell walls of terrestrial plants. Isolated pectin, which is rich in galacturonic acid, has a typical molecular weight of 60,000–100,000 Da, varying according to the extraction conditions and origin.

Pectin hydrogels are useful chip surfaces with low nonspecific background for the analysis of biomolecules expressed in plants.

Due to the abundant carboxyl groups, PC coatings are fully compatible with all immobilization chemistries available for CM-Dextran sensor chips.

Polyethylene glycol (PEG)

The PEG sensor surface coating is a hydrophilic, polar surface based on a dense brush of 4-kDa PEG. The low polymer–water interfacial energy value results in good resistance to protein adsorption and cell adhesion.

The PEG surface is hydroxyl terminated and uncharged. It can be activated by oxidation to aldehydes and subsequent Schiff base coupling/reductive amination, or by (bis)epoxy activation. As there will not be any electrostatic preconcentration, high ligand concentrations are necessary to reach a sufficient level of immobilization.

Due to the terminal functionalization of the PEG chains, this surface is useful for the preparation of ligand monolayers on a well-stabilized, semi-hydrated and passive spacer layer.

Poly-L-lysine (PLY)

Pure poly-L-lysine brush, 50–100 nm thick. High positive charge density. Sensor chips coated with PLY can be used for reverse electrostatic preconcentration and immobilization of amino reactive, negatively-charged ligands, polymers or biological structures, such as cells and tissue fragments. Note that the nonspecific background of this surface can be significant, as most biomolecules carry a negative charge at physiological pH.

Streptavidin derivatized carboxymethyldextran (SAD)

The streptavidin-modified carboxymethyl dextran matrix consists of covalently bound streptavidin in a biocompatible, hydrophilic polysaccharide. The 55-kDa streptavidin tetramer is composed of four identical polypeptide chains and is dissociated into monomers before ligand immobilization, which prevents baseline drift and leaching during the ligand regeneration steps.

Due to the mechanical rigidity of the polymer backbone, combined with branching and pronounced hydration, the movement of immobilized ligands in SAD polysaccharide hydrogels is limited. Therefore, SAD hydrogels are a good regime if the focus is on monovalent interactions. If immobilized ligands are expected to interact in a cooperative manner, the more flexible SAHC hydrogel should be used instead.

Streptavidin derivatized HC polycarboxylate (SAHC)

The streptavidin-modified HC matrix consists of covalently bound streptavidin in a bioinert, extremely hydrated, strictly linear polycarboxylate. The 55-kDa streptavidin tetramer is composed of four identical polypeptide chains and is dissociated into monomers before ligand immobilization, which prevents baseline drift and leaching during the ligand regeneration steps.

SAHC hydrogels exhibit a very low nonspecific background. The small molecular footprint of the HC polymer chains leaves sufficient space for unhindered diffusion of the analyte, even after immobilization of voluminous ligands. The polymer chains of the SAHC hydrogel are extremely flexible, giving immobilized ligands a high degree of freedom and the ability to interact in a cooperative manner with large analytes.

Streptavidin immobilized on 2D carboxymethyldextran (SAP)

The streptavidin-modified planar (two-dimensional, 2D) carboxymethyldextran matrix consists of covalently-bound streptavidin on a thin (<5 nm), biocompatible, hydrophilic polysaccharide cushion. The 55-kDa streptavidin tetramer is composed of identical polypeptide chains that are dissociated into monomers before ligand immobilization to prevent baseline drift and leaching during the ligand regeneration steps.

Because of the planar nature of the SAP structure, the immobilization capacity is significantly lower than that of 3D streptavidin coatings (LINK SAD, SAHC). However, the planar variant has significant advantages over the 3D hydrogels in the kinetic measurement of larger analytes such as antibodies. Because of the absence of flexible polymer strands, the immobilized ligands are spatially resolved on the surface and mainly monovalent. Furthermore, analytes on the planar SAP structure can diffuse unhindered to, and dissociate from, the surface without steric hindrance caused by 3D matrices. Moreover, analyte re-binding during the dissociation phase does not occur. All these factors mean that kinetic data obtained in measurements on the SAP surface are more reliable than comparable measurements made using 3D hydrogels.

Tetraethylene glycol (TEG)

Tetraethylene glycol (TEG) is a member of a homologous series of dihydroxy alcohols and is well known for its biocompatibility. The sensor chip coating consists of a TEG self-assembled monolayer (SAM), which exclusively presents hydroxyl groups on the surface. Sensor chips with TEG SAMs have the advantage that the interactions take place very close to the gold surface, and thus in the most sensitive part of the evanescent field, resulting in acceptable SPR signals with medium-weight molecules as analytes. The TEG surface is ideal for the development of biocompatible surfaces, and can be used as a reference because of its well-documented ability to suppress protein adsorption and plasma activation. Furthermore, TEG can be modified chemically to alter the properties of the surface1, 2. Use of modified TEG sensor chips is of interest in the pharmaceutical and cosmetics industries, as TEG is often used as an additive in their products.

For TEG-based sensor chips with some carboxyl groups on the surface, which allow easy and gentle immobilization via EDC/NHS chemistry, please select CMTEG sensor chips.

Cha, T., Guo, A., Jun, Y., Pei, D., & Zhu, X. Y. (2004). Immobilization of oriented protein molecules on poly (ethylene glycol)‐coated Si (111). Proteomics, 4, 1965–1976.
Piehler, J., Brecht, A., Valiokas, R., Liedberg, B., & Gauglitz, G. (2000). A high-density poly (ethylene glycol) polymer brush for immobilization on glass-type surfaces. Biosensors and Bioelectronics, 15, 473–481.

Disulfide modified hydrogels (THC, TD, TP)

THC, TD and TP sensor chips are coated with a disulfide derivatized bioinert hydrogel respectively 2D coating based either on carboxymethyldextran or the linear polycarboxylate HC. Covalent disulfide coupling requires reduction of surface and ligand disulfides and activation of the formed surface thiol groups with pyridyl disulfide before ligand injection. All THC, TD and TP coatings also contain carboxyl groups for electrostatic preconcentration of protein ligands.

UV-reactive sensor chip surfaces

The UV crosslinking surface makes immobilization very easy. Let your molecule of interest preconcentrate on the surface, then irradiate with near-UV light. A high immobilization yield and an immobilization chemistry that targets many functional groups such as hydroxyls and amines, as well as less reactive C–H and C–C bonds, allow the immobilization of carbohydrates and small molecules which are not accessible through traditional NHS/amine coupling. The wavelength and UV dose required for good coupling yields do not affect even sensitive ligands.

The UV chemistry is compatible with the Reichert 2SPR system equipped with a UV flow cell, and SPRi instruments using external spotters. Alternatively, it is possible to undock a sensor chip after ligand preconcentration, irradiate the surface with UV, and reinsert the chip to continue the experiment.

Zwitterionic surfaces (ZC, ZCX, ZCC)

This unique hydrogel is a derivative of the robust C polycarboxylate. It has been developed for covalent immobilization of negatively charged molecules such as DNA, carbohydrates and acidic proteins at physiological pH, employing a reverse electrostatic preconcentration effect. In contrast with the usually polyanionic surfaces for covalent immobilization, this surface exhibits a temporarily positive net charge during preconcentration and coupling of the negatively charged ligand. After coupling and quenching of the active NHS esters, the net charge becomes slightly negative again, thus stabilizing the surface against nonspecific interactions.

The zwitterionic hydrogel is available as an NHS-preactivated surface (ZCX) or a non-activated surface (ZC) to be activated in situ with standard EDC/NHS chemistry.

To determine preconcentration conditions(preconcentration scouting), we offer the positively charged ZCC version, which mimics the NHS activated surface. However, the ZCC surface is unreactive, so the ligand can be eluted multiple times until optimization of the coupling parameters has been completed.

Products | SPR Sensor chips

SPR Sensor chips

XanTec bioanalytics provides laboratories worldwide with the ultimate in SPR biosensor chips via superior coating technologies. Our proprietary technology, developed over more than 25 years ago, solves a number of critical issues usually associated with state of the art Self Assembled Monolayer (SAM) surface chemistry. The coatings are robust and prevent exposure of hydrophobic nanodomains or pinhole defects which can cause non-specific interactions. As the chemistry is extremely versatile, a large choice of topcoats is available to specifically address different experimental needs.

Not only do we feature the largest portfolio of sensorchip coatings and chemistries in the world, we offer the versatility to work on a number of different instruments and in varying formats. The chips fit into instruments of all major manufacturers. In addition, we provide customized sizes or can coat alternative substrate materials.

Short brochure SPR chips

Compatible sensor chips and prisms
Biacore™
BioNavis™
Biosensing Instrument
Bruker™ / Sierra Sensors™
Sartorius™ / ForteBio™ / SensiQ™
Reichert SPR
BioRad™ ProteOn®
IBIS MX96®
IBIS/Kinetic Evaluation Instruments™
Horiba™

Instrument compatibility of XanTec's SPR Sensor chips/prisms. Other manufacturers are covered by OEM contracts.

Biacore™ users can find the XanTec sensor chip corresponding to their Biacore™ sensor chip in the following table.

Modification	Cytiva™ sensor chips	XanTec equivalent	Xantec alternative
Plain gold surface	Au	AU
No hydrogel, low capacity	C1	CMDP planar	HCP planar
Short matrix, normal capacity	CM3	CM50L	HC30M
Standard hydrogel, low charge density	CM4	HLC200M	HLC30M
Standard hydrogel, normal capacity	CM5	CMD200M	CMD200L, HC200M
Standard hydrogel, high capacity	CM7	CMD700L	CMD500M, HC1500M
Biotin capturing surface	SA	SAD200M	SAD200L, SAHC200M
PEG-based alternative to dextran surface	PEG	CMPEG	CMTEG
Hydrophobic 2D surface	HPA	HPP
Vesicle and liposome capture	L1	LD	LP
NTA for His6 tagged ligands	NTA	NiD200M	Poly-NTA NiHC group (high affinity)
Antibody capture surface	Protein A	PAD200L	PAHC200M
Antibody capture surface	Protein G	PAGD200L	PAGHC200M
Reversible capture of biotinylated ligands	CAPture Kit	RGD200M

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