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 can generate 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

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 and 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 the high affinity and specificity observed for rabbit antibodies. Phage display normally does not support such modifications and consequently 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 range of 0.001-1.0 nM as demonstrated by a recently completed contract project for the National Cancer Institute (NCI)[HHSN261201300023C].

An example of a rabbit monoclonal antibody clone generated for this project with AvantGen’s technology that can specifically distinguish a peptide in its phosphorylated and non-phosphorylated form is shown below together with a rabbit monoclonal antibody clones against polysaccharides from group A Streptococcus (Strep A).

Phospho-specific rabbit monoclonal antibody

A rabbit monoclonal antibody clone against a phosphorylated peptide derived from a cancer antigen, AVG309: NH2-RPHFPQF-pS-YSASGTA-OH (phosphopeptide). This clone has a KD=0.02 nM 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 than the KD value.

A rabbit monoclonal antibody clone against a phosphorylated peptide derived from a cancer antigen, AVG309: NH2-RPHFPQF-pS-YSASGTA-OH (phosphopeptide). This clone has a KD=0.02 nM 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 than the KD value.

Bacterial strain (Strep A)-specific rabbit monoclonal antibody

A rabbit monoclonal antibody clone against Strep A, that specifically recognizes different strains of Strep A, but not other strains, including Strep-B, C, F or G. The affinity for this clone, H2, is 0.05 nM

A rabbit monoclonal antibody clone against Strep A, that specifically recognizes different strains of Strep A, but not other strains, including Strep-B, C, F or G. The affinity for this clone, H2, is 0.05 nM

 

From antigen to rabbit monoclonal antibody-producing cells in less than 5 months

diagram

 

Harness the advantages of rabbit monoclonal antibody for your research needs

For novel epitope recognition
For differentiation between subtle structural variations
For detection of post-translational modifications
For IHC study 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
Experience gained from working with 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 contact us: service@nullavantgen.com