Jackson Immuno Research Inc.
specializing in secondary antibodies and conjugates

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Secondary Antibodies for VHH Discovery

Secondary Antibodies for VHH Discovery

JIR presents Anti-Alpaca IgG, Subclass and VHH domain secondary antibodies to optimize VHH (single domain) antibody (nanobody*) development and application.

VHH Antibody Development Process

The pathway to recombinant VHH antibody development

The development of high affinity VHH antibodies follows the well-established pathway shown above.

JIR Anti-Alpaca antibodies can be utilized at specific stages to expedite and enhance the development of high affinity VHH antibodies. The success and speed of discovering high quality VHH candidates is optimized by the fidelity and utility of the tools used at each step.

Browse Anti-Alpaca IgG, Subclass and VHH domain Secondary Antibodies


Monitor IgG 2+3 subclass titer to ideally time PBMC harvest

Monitor Camelid IgG Subclasses

Use JIR Anti-Alpaca IgG, subclasses 2+3 to time PBMC harvest for VHH library construction. The VHH domain is located on these subclasses only. Subclass 2+3 response may mature differently from total IgG, and monitoring serum conversion of these antibodies can inform the decision to harvest PBMCs. Timing PBMC harvest based on total IgG titer might suggest a collection point at 150 days, but the IgG 2+3 response was not maximal until after 300 days post immunization. This ELISA data shows the antibody subclass repertoire in response to the immunogen over time. Antiserum was used to detect the coated immunogen, and peroxidase-conjugated secondary antibodies were used to compare total IgG to IgG subclasses 2+3.

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Enhance detection of VHH antibodies using Goat Anti-Alpaca IgG, VHH domain

VHH antibody detection by Western blot

Anti-Alpaca IgG, VHH domain antibodies show amplified signal compared to an anti-tag antibody. Polyclonal antibodies that broadly recognize many epitopes on VHH deliver additional signaling molecules (e.g. fluorophores or enzymes) vs. antibodies narrowly focused on small peptide sequences. In this figure, a His6 tagged recombinant VHH was detected by western blot using Goat Anti-VHH domain antibody or an anti-His6 antibody.


Test for VHH expression and binding activity with greater confidence 

Test for VHH antibody expression with JIR Anti-Alpaca IgG, VHH domain

Screening VHH-expressing clones for solubility and binding activity is commonly done by ELISA. Significant signal can be obtained even at low VHH concentrations. In this panel, VHH was titrated across its coated immunogen and detected with Peroxidase conjugated Goat Anti-Alpaca VHH domain antibody.


Anti-Alpaca IgG Secondary Antibodies

Secondary Antibody

Code

Goat Anti-Alpaca IgG (H+L)**

128-005-003

Goat Anti-Alpaca IgG (H+L) (min X Bov, Hu, Ms, Rb, Rat Sr Prot)**

128-005-160

Goat Anti-Alpaca IgG, subclasses 2+3 specific (min X Bov, Hu, Ms, Rb, Rat Sr Prot)**      

128-005-229

Goat Anti-Alpaca IgG, VHH domain (min X Bov Sr Prot)

128-005-232

Goat Anti-Alpaca IgG, VHH domain (min X Bov, Hu, Ms, Rb, Rat Sr Prot)

128-005-230

JIR’s Anti-Alpaca antibodies recognize both alpaca and llama IgG (H+L), subclasses 2+3, or VHH domain.

**These antibodies react primarily with the Fc region, and are not recommended for detection of VHH antibodies.

Conjugate Options

  • Unconjugated
  • Horseradish Peroxidase
  • Alkaline Phosphatase
  • Biotin-SP
  • DyLight™ 405
  • Alexa Fluor® 488
  • FITC
  • Cy™ 3
  • R-Phycoerythrin
  • Alexa Fluor® 594
  • Rhodamine Red-X™
  • Alexa Fluor® 647
  • Cy™ 5

AffiniPure™ Antibodies is trademark of Jackson ImmunoResearch Laboratories, Inc. 

*Nanobody is a registered trademark of Ablynx N.V

DyLight™ fluorescent dyes is a trademark of Thermo Fisher Scientific. 

Cy™ is a registered trademark of GE Healthcare. 

Rhodamine Red™-X is a trademark of Invitrogen. 

Alexa Fluor® is a trademark of Life Technologies Corp.

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