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Nano-CaptureLigand human IgG

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BLI binding kinetics of a human IgG1 anti-IgE antibody to human IgE. A human IgG1 anti-IgE 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 human IgE.
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).

Description
The Nano-CaptureLigand™ human IgG captures and immobilizes the Fc-fragment of human IgG antibodies to avidin/streptavidin coated surfaces used in BLI, SPR, and ELISA assays. It consists of a biotinylated Nanobody that specifically binds to human IgG antibodies with high affinity and in a site-directed manner.

Specificity: Fc-fragment of human IgG1, IgG2, IgG3, and IgG4; Co-reactive to rabbit IgG
Nano-CaptureLigands specificity overview

Conjugates: Biotin

Applications
Human IgG capture to biosensors used in BLI and SPR
Human IgG immobilization for ELISA

Advanced immobilization of antibodies
Specific, stable, and site-directed immobilization of non-biotinylated human IgG antibodies to streptavidin/avidin

 

Product Size Code Price Buy
Product Nano-CaptureLigand™ human IgG/rabbit IgG, Fc-specific VHH, biotinylated Size 10 µL Code shurbGB-1-10 Price $ 130
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Product Nano-CaptureLigand™ human IgG/rabbit IgG, Fc-specific VHH, biotinylated Size 100 µL Code shurbGB-1-100 Price $ 480
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Product Type
Capture Nanobody (VHH)

Applications
Immobilization of human antibodies on avidin and streptavidin surfaces for Bio-Layer Interferometry (BLI), Surface Plasmon Resonance (SPR), and ELISA

Target/ Specificity 
Fc-fragment of human IgG1, IgG2, IgG3, and IgG4
Co-reactive to Fc-fragment of rabbit IgG

Cross-reactivity
No cross-reactivity to human IgA, IgE, IgM; mouse IgG1, IgG2a, Ig2b, IG2c, IgG3; rat IgG1, IgG2a, IgG2b, IgG2c; sera from goat, guinea pig, rat, sheep
Cross-reactivity to macaque (cynomolgus monkey) serum

Affinity (Kd) of monovalent (1:1) binding mode
Human IgG1: 1.4 nM; Human IgG2: 0.4 nM; Human IgG3: 9.0 nM; Human IgG4: 0.2 nM
Apparent affinity may be higher for full IgGs due to avidity effects (1 antibody captured by 2 Nanobodies).

Concentration 
1 g/L (71 μM)

Conjugate
Biotin

Degree of biotinylation 
On average 1-2 biotin molecules per Nanobody

Format 
Alpaca single domain antibody, monovalent

Host
Alpaca-derived, recombinantly produced in bacteria

Clonality
Monoclonal

Clone
CTK0101 (VHH0238)

How to cite this product
Nano-CaptureLigand™ human IgG/ rabbit IgG, Fc-specific VHH, biotinylated
RRID: AB_2848185

Synonyms 
Alpaca single domain antibody, VHH, Nanobody, binding domain of single domain antibody, Nano-antibody

Validation 
Application validated for ELISA and BLI (FortéBio Octet® systems)
Determination of cross-reactivity, subclass specificity, sequence, affinity, and melting temperature

Purity
Recombinantly expressed and purified via His-tag

Form
Buffered aqueous solution

Storage buffer 
25 mM TAPS pH 8.5, 500 mM NaCl, 5 mM EDTA, Preservative: 0.09 % sodium azide

Storage conditions 
Upon receipt store at +4°C/+40°F. Optional: Aliquot upon arrival and store at -20°C/-4°F

Stability
Stable for 1 year at +4°C/+40°F

Shipment 
Shipped at ambient temperature

Applications

  • Detection and quantitation of human IgG antibodies and their antigens
  • Characterization of human IgG antibodies’ binding kinetics and affinity
  • Human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma
  • Epitope binning

The next level of antibody immobilization to BLI and SPR biosensors

  • Site-directed and specific immobilization of the Fc-fragment of human IgG to streptavidin coated biosensors without biotinylation of the antibody
  • Compatible to surface plasmon resonance (SPR) and biolayer interferometry (BLI) assays (e.g. FortéBio Octet® systems)
  • High affinity for stable baseline without significant dissociation
  • >10x regeneration for multiple reuse
  • High specificity enables human IgG antibody discovery and screening in crude liquids like hybridoma supernatant, serum, and plasma samples

 

Comparison of BLI/SPR immobilization methods

The next level of antibody immobilization for ELISA

  • Compatible to enzyme-linked immunosorbent assays (ELISA) 
  • Site-directed immobilization of Fc-fragment of human IgG antibodies to streptavidin/avidin coated surfaces of sandwich ELISAs without biotinylation of antibody
  • Combine the site-specificity of protein A, G, and L with the stability of the biotin/streptavidin interaction
  • Human IgG specificity allows the application of a broad range of detection antibodies 
  • High affinity for strong immobilization of human IgG antibodies even at low antibody concentrations

 

Comparison of ELISA immobilization methods

Nano-CaptureLigands are comprehensively validated

  • Characterized: DNA and protein sequences, affinity, melting temperature
  • Recombinantly produced, virtually without lot-to-lot variation
  • Manufacturing process is animal free

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Only for research applications, not for diagnostic or therapeutic use!