FAQs on Rabbit Monoclonal Antibody Development
Scientist
on
May 14, 2024
This blog post was written by AvantGen, a pioneering biotechnology CRO dedicated to accelerating antibody-based therapeutics as well as providing critical reagent and diagnostic antibodies for the research community. In this blog post, AvantGen answers frequently asked questions about its longest continually offered service: rabbit monoclonal antibody development. For more information about AvantGen and its innovative work, please visit www.avantgen.com.
1. Why use rabbit antibodies instead of mouse antibodies?
Rabbit antibodies offer several advantages over mouse antibodies. Firstly, rabbits produce a more diverse antibody repertoire due to their larger immune system and the ability to undergo gene conversion, leading to antibodies with much higher specificity and affinity. Additionally, rabbit antibodies recognize epitopes that may not be immunogenic in mice, making them suitable for targets that are challenging to raise antibodies against in mice. Furthermore, the evolutionary distance between rabbits and humans results in rabbit antibodies that often exhibit superior cross-reactivity to human antigens, making them valuable tools for research and diagnostics. This may in part be due to the fact that most mouse strains used are inbred, which in general elicit less diverse immune responses. A head-to-head comparison of rabbit versus mouse derived antibodies used in immunohistochemistry (IHC) staining demonstrated consistently higher sensitivity from rabbit mAbs.1
2. What are the applications of rabbit monoclonal antibodies?
Rabbit monoclonal antibodies have a wide range of applications across various fields of research and diagnostics. They are commonly used in techniques such as immunohistochemistry, Western blotting, flow cytometry, immunoprecipitation, ELISA and ChIP assays. Additionally, they are often used for detecting small and large molecule drugs in pharmacokinetic studies or as a critical step in the manufacturing and quality control of biologics.
3. How are rabbit monoclonal antibodies generated?
Rabbit monoclonal antibodies against a specific target are typically generated in a two-step process: immunization and antibody screening. Rabbits are immunized with the specific target in the form of a small molecule, peptide, protein, DNA/RNA or cells. Adjuvants, conjugates and/or carriers are often used to stimulate the animal’s antibody response. Animals typically receive subsequent boosters to further elicit a robust immune response. Serum drawn from the animals can be measured for target-specific antibodies and can be used to determine the appropriate time to collect antibody secreting B cells for screening. Multiple methods are available for screening, including hybridoma, single B-cell and phage and yeast display technologies to identify specific monoclonal antibodies. The success of screening, especially in identifying rare monoclonal antibodies that bind to a unique epitope, often comes down to two things: the size of the haystack and the ability to find that needle. With speed and cost in mind, different technologies have varying efficiencies.
4. Where can I find high-quality rabbit antibodies for my research?
High-quality rabbit antibodies can be obtained from reputable commercial suppliers. A handful of contract research organizations (CROs) in particular have expertise in generating rabbit monoclonal antibodies against custom or novel targets. AvantGen has a long history of providing high-quality rabbit mAbs to the research community, including therapeutic, diagnostic, nonprofit and government entities for various applications. By combining proprietary immunization protocols with a novel and robust yeast display system, one in a million ultra-specific antibodies can be identified, often discriminating between a single amino acid, post-translational modification or chemical moiety in the target molecule. AvantGen also produces rabbit monoclonal antibodies in recombinant form with known sequences. Furthermore, the potential FcRn binding activity of rabbit Fc domain can be eliminated to reduce background binding activity in various cell-based assays, such as IHC and flow cytometry. To learn more about AvantGen’s AvantBunny platform and services, please contact partner@avantgen.com or connect via the Scientist.com link below.
5. And lastly, any recommendations for a rabbit children’s book?
The Velveteen Rabbit by Margery Williams. For children and adults alike. Enjoy!
References
- Weber J, Peng H, Rader C. From rabbit antibody repertoires to rabbit monoclonal antibodies. Exp Mol Med. 2017 Mar 24;49(3). PMID: 28336958.