Rabbit monoclonal antibodies are impressive and efficient tools, from analytical applications (e.g., ELISA, western blot, and Immunostaining) to diagnostics (e.g., detection of tumor-related antigens: PDL-1, HER2, and estrogen receptor) and as therapeutics also (e.g., Eptinezumab, Brolucizumab).
Compared to mouse, rabbit monoclonal antibodies have higher affinity and specificity for their targets.
Pei Liu, Ph.D., Senior Scientist, Antibody Department, GenScript
Pei Liu Ph.D., Senior Scientist, Antibody Department, GenScript. Image Credit: GenScript.
For therapeutic use, the higher sensitivity and epitope specificity of rabbit monoclonal antibodies translate to reduced off-target binding and lower dosage requirements, ultimately minimizing the potential for toxicity.
Broader antigen detection capabilities
Rabbit monoclonal antibodies provide more extensive target recognition capacities that are suitable for therapeutics and diagnostics. Broader paratope diversity is shown by the unique processes that give rise to the primary antibody repertoire in rabbits, which tends to be dependant on somatic hypermutations, as in mice, but also on somatic gene conversion, a type of homologous recombination that is predominantly found in chicken and rabbit.
Expanded opportunities for screening
The larger size of a rabbit, particularly its larger spleen and higher blood volume and bone marrow tissue, implies better chances for detaching B cells and looking for more monoclonal antibodies with the required epitope specificities.
Cross-reactivity is a plus
The cross-reactivity of rabbit-derived monoclonal antibodies with epitopes in human and mouse antigens is an added benefit. This is useful in preclinical verification and depiction of human disease mouse models.
The rabbit’s evolutionary distance from human is further than that of rodents, allowing the rabbit immune system to generate monoclonal antibodies against human antigens commonly non-immunogenic in mice.
Sasidhar Murikinati, Ph.D., Regional Marketing Manager, ProBio, GenScript
Sasidhar Murikinati Ph.D., Regional Marketing Manager, ProBio, GenScript. Image Credit: GenScript.
Low immunogenicity antigens are not a problem
The immune system of rabbits can identify human antigen epitopes, which are otherwise neglected by rodents.
If the target in discussion tends to have low immunogenicity, which is archetypal for tiny molecules, rabbits could be the ideal option for monoclonal antibody development. Contrary to rodents, rabbits can mount a tougher immune response from haptens, tiny molecules, and post-translational modifications (e.g., AMPylated peptides and phosphorylated peptides).
Commercially available polyclonal antibodies to detect AMPylated proteins were not specific enough to detect AMPylated proteins in complex samples for target identification. Anti-AMP monoclonal antibodies were developed because of their potential as an orthogonal method that may be broadly used by any lab.
Dorothea Hopfner, Ph.D. Candidate, Department of Chemistry, Technical University Munich
Dorothea Hopfner, Ph.D. Candidate, Department of Chemistry, Technical University Munich. Image Credit: GenScript.
According to Dorothea Hopfner, when developing monoclonal antibodies against post-translational modifications, researchers should:
- Identify small moieties with great affinity
- Identify small but significant differences among various post-translational modifications (e.g., di- and tri-methylation)
- Finally, if the antibody is needed to be utilized for new target recognition, a desirable characteristic is that the antibody can identify post-translational modifications without the dependence on protein sequence
Synthetic peptides are an asset in producing anti-PTM antibodies. “Synthetic peptides can significantly help in anti-PTM antibody development, and the bottleneck of antibody candidate selection can be overcome by including native modified protein for screening when immunizing with peptides,” stated Dorothea Hopfner.
GenScript generated an antibody using a peptide of phosphorylated BRCA1 and this was a very useful tool. The anti-BRCA1 (pS114) rabbit polyclonal antibody reagent was specific, evidenced by the lack of signal with unphosphorylated BRCA1.
Jo Morris, Ph.D., Professor, Molecular Genetics, Institute of Cancer and Genomic Sciences, University of Birmingham
Jo Morris Ph.D., Professor, Molecular Genetics, Institute of Cancer and Genomic Sciences, University of Birmingham. Image Credit: GenScript.
Altogether, the improved characteristics of rabbit antibodies are of great benefit for applications that require great sensitivity and epitope specificity. Whether a rabbit monoclonal or polyclonal antibody is selected ultimately depends on the user's individual requirements.
For the detection of the AMPylated protein, Dorothea Hopfner needed a long-term, reliable, and consistent antibody source to upkeep characterization and authentication efforts. Thus, a rabbit monoclonal antibody was the ideal option to prevent issues associated with batch-to-batch reproducibility.
Genscript is the world’s leading biotech company providing life sciences services and products. With gene synthesis, peptide, protein, antibody and preclinical drug development service capabilities, we are internationally recognized as a leading biotech company specializing in fundamental life sciences research and early-phase drug discovery services. As of 2018, more than 30,000 peer-reviewed journal articles cited GenScript’s services and products, making GenScript the most frequently cited biotech company in the world.
After almost two decades of fast growth in developing biological reagents, the company has expanded its business into immunotherapy, CDMO, laboratory equipment, and microbial industry to further fulfill its mission in making people and nature healthier through biotechnology.
Founded in 2002 in New Jersey, United States, GenScript serves as a partner for researchers in basic life sciences, translational and biomedical fields as well as early-stage drug development.
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