Novel Rabbit Monoclonal Antibody Generation
Superior Affinity and Specificity and Antibody Clones Characterized at the DNA Sequence Level

 

Advantages of rabbit monoclonal antibodies

The rabbit immune system generates and affinity matures antibodies by mechanisms that differ from those of mice and other rodents. Antibodies produced by rabbits normally have 10 to 100-fold higher affinity for antigen than mouse monoclonal antibodies. In addition, the rabbit immune system is particularly adept at generating antibodies that are able to distinguish between very similar molecules with subtle structural variations.

​Advantages of AvantGen’s technology for generation of rabbit monoclonal antibodies

AvantGen’s yeast display system is well suited for displaying rabbit antibodies whose light chains normally have an intra-light chain disulfide bond that is required for maintaining their high affinity and specificity.

By combining an optimized rabbit immunization protocol and AvantGen’s robust yeast display system, we are able to generate a yeast display library of sufficient size to isolate very high affinity monoclonal antibodies produced by immunized rabbits. The yeast display system is uniquely suited for displaying rabbit antibodies, which have an additional disulfide bond in their light chain. This disulfide bond formation is believed to be important for structural stability and supports the high affinity and specificity observed for rabbit antibodies. Phage display normally does not support such modifications, and therefore is not well suited for displaying rabbit antibodies for screening.

Our approach overcomes issues associated with rabbit hybridoma-based technology, such as low fusion efficiency, unstable cell lines and low antibody yield, and greatly shortens the time required to generate monoclonal antibody producing stable cells. Antibodies generated with our technology often exhibit affinities in the low pM range as demonstrated by a recently completed contract project for the National Cancer Institute (NCI) [HHSN261201300023C].

The following is an example of rabbit monoclonal antibodies generated for the NCI’s project with AvantGen’s technology that can specifically distinguish a peptide in its phosphorylated and non-phosphorylated form and another example of rabbit monoclonal antibody against polysaccharides from group A Streptococcus (Strep A).

Phospho-specific rabbit monoclonal antibody

Rabbit monoclonal antibody against a phosphorylated peptide derived from a cancer antigen, AVG309: NH2-RPHFPQF-pS-YSASGTA-OH (phosphopeptide) with a KD=20pM and does not recognize its non-phosphorylated counterpart (AVG311 NH2-RPHFPQFSYSASGTA-OH) or other unrelated peptides (AVG301-308) even at 1000 fold higher antibody concentration

Bacterial strain (Strep A)-specific rabbit monoclonal antibody

Rabbit monoclonal antibody against Strep A, specifically recognizes different subtypes of Strep A, but not other strains, including strep-B, C, F or G. The affinity for this clone, H2, is 50pM

A typical procedure and timeline for generation of rabbit monoclonal antibodies is illustrated below:

diagram

 

Harness the advantages of rabbit monoclonal antibody for your research and diagnostic needs

  • For novel epitope recognition
  • For differentiation between subtle structural variations
  • For detection of post-translational modifications
  • For IHC studies with low background
  • For use of mouse proteins and tissues

AvantGen’s custom services offer you:

  • High-quality rabbit monoclonal antibodies tailored to your needs
  • Dedicated team assigned to your project
  • Dedicated and experienced team assigned to various targets from the NIH and commercial partners
  • Cost and time saving benefits for your project
  • Rabbit mAb-expressing stable cell lines
  • Monoclonal antibody clones characterized at the DNA sequence level for consistent batch-to-batch performance

For more information and inquiries about AvantGen’s rabbit monoclonal antibody services, please complete the request form and send it to service@nullavantgen.com