Structure of VHH.

SDS-PAGE of EGFP: 10 % acrylamide gel; 1 µg of EGFP has been loaded. Detection: InstantBlue

Normalized EGFP fluorescence excitation and emission spectra.

BLI binding kinetics of a human anti-GFP Fab fragment to eGFP. A synthetic human anti-GFP lambda Fab fragment was immobilized using Nano-CaptureLigand human Ig, lambda-LC-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of eGFP (ChromoTek EGFP, recombinant protein).

Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of lambda light chain of human Ig antibody (orange). The setup is ready for characterization of human IgG antibodies’ binding kinetics and affinity to substrate (red), human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

ELISA capture of a human IgG1 lambda antibody using Nano-CaptureLigand human Ig, lambda-LC-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human Ig, lambda-LC-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Human IgG1 lambda antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

Species reactivity of Secondary VHH.

BLI binding kinetics of a human IgE antibody. 20 nM human IgE antibody was immobilized using Nano-CaptureLigand human IgE, VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. Human IgE is stably captured by the Nano-CaptureLigand with negligible dissociation.

BLI binding kinetics of human, mouse and rabbit antibodies. 20 nM of human, mouse and rabbit antibodies were immobilized using Nano-CaptureLigand human IgE, VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. Only human IgE antibody is captured by the Nano-CaptureLigand with negligible dissociation.

Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) assay followed by capture of an antibody (orange). The setup is ready for characterization of the antibodies’ binding kinetics and affinity to target (red), antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

ELISA capture of a human IgE antibody using Nano-CaptureLigand human IgE, VHH, biotinylated. 50 nM Nano-CaptureLigand human IgE, VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Human IgE antibody was titrated in a 1:2 dilution series and detected with a rabbit anti-human IgE primary antibody and an alkaline phosphatase-conjugated detection antibody.

Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

Species reactivity of Secondary VHH.

BLI binding kinetics of a monoclonal rabbit IgG anti-DYKDDDDK-tag antibody to a DYKDDDDK-tagged protein. A monoclonal rabbit IgG anti-DYKDDDDK-tag antibody was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a DYKDDDDK-tagged protein.

ELISA capture of a rabbit IgG antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Rabbit IgG antibody were titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

BLI binding kinetics of a polyclonal rabbit IgG anti-GFP antibody to eGFP. A polyclonal rabbit IgG anti-GFP antibody (ChromoTek GFP antibody [PABG1]) was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of eGFP (ChromoTek EGFP, recombinant protein).

BLI binding kinetics of different human IgG antibodies. Different monoclonal human IgG antibodies were immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. All four human IgG subclasses are stably captured by the Nano-CaptureLigand with negligible dissociation.

Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of human IgG antibody (orange). The setup is ready for characterization of human IgG antibodies’ binding kinetics and affinity to substrate (red), human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

ELISA capture of human IgG1 lambda antibody using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Human IgG1 lambda antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

Species reactivity of Secondary VHH.

BLI binding kinetics of a mouse IgE antibody. 20 nM mouse IgE antibody was immobilized using Nano-CaptureLigand mouse IgE, VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. Mouse IgE is stably captured by the Nano-CaptureLigand with negligible dissociation.

ELISA capture of mouse IgE antibody using Nano-CaptureLigand mouse IgE, VHH, biotinylated. 50 nM Nano-CaptureLigand mouse IgE, VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Mouse IgE antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) assay followed by capture of an antibody (orange). The setup is ready for characterization of the antibodies’ binding kinetics and affinity to target (red), antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

BLI binding kinetics of mouse, human and rabbit antibodies. 20 nM of mouse, human and rabbit antibodies were immobilized using Nano-CaptureLigand mouse IgE, VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors. Only mouse IgE antibody is captured by the Nano-CaptureLigand with negligible dissociation.

Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

Species reactivity of Secondary VHH.

BLI binding kinetics of a mouse IgG1 anti-DYKDDDDK-tag antibody to a DYKDDDDK-tagged protein. A monoclonal mouse IgG1 anti-DYKDDDDK-tag antibody was immobilized using Nano-CaptureLigand mouse IgG1, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a DYKDDDDK-tagged protein.

BLI binding kinetics of a monoclonal rabbit IgG anti-DYKDDDDK-tag antibody to a DYKDDDDK-tagged protein. A monoclonal rabbit IgG anti-DYKDDDDK-tag antibody was immobilized using Nano-CaptureLigand human IgG/rabbit IgG, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a DYKDDDDK-tagged protein.

BLI binding kinetics of a purified and a hybridoma supernatant mouse IgG1 anti-Halo antibody to Halo-tag. A purified, monoclonal mouse IgG1 anti-Halo antibody (blue, ChromoTek Halo antibody [28A8]) and a hybridoma supernatant, monoclonal mouse IgG1 anti-Halo antibody (green) were immobilized using Nano-CaptureLigand mouse IgG1, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of Halo-tag protein.

ELISA capture of mouse IgG1 antibody using Nano-CaptureLigand mouse IgG1, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand mouse IgG1, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Mouse IgG1 antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

Species reactivity of Secondary VHH.

BLI binding kinetics of a mouse IgG2a anti-V5-tag antibody to a V5-tagged protein. A monoclonal mouse IgG2a anti-V5-tag antibody was immobilized using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated on FortéBio Streptavidin (SA) Biosensors and assayed with different concentrations of a V5-tagged protein.

Immobilization of the Nano-CaptureLigand (green) on Biosensor (white) for biolayer interferometry (BLI) followed by capture of mouse IgG2a antibody (orange). The setup is ready for characterization of mouse IgG2a antibodies’ binding kinetics and affinity to substrate (red), mouse IgG2a antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma.

ELISA capture of mouse IgG2a antibody using Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated. 50 nM Nano-CaptureLigand mouse IgG2a, Fc-specific VHH, biotinylated was used for coating on an avidin-coated MaxiSorp plate. Mouse IgG2a antibody was titrated in a 1:2 dilution series and detected with an alkaline phosphatase-conjugated detection antibody.

Sandwich ELISA: Immobilization of the Nano-CaptureLigand on streptavidin coated microplates for ELISA. No biotinylation of the antibody is required. Coating of the streptavidin microplate with Nano-CaptureLigand (top left). Site-directed antibody immobilization via Nano-CaptureLigand (top right). Addition of sample including target (red, bottom left). Addition of detection and secondary antibody for read-out (bottom right).

Species reactivity of Secondary VHH.