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Rabbit immunization + Fab-phage display
Why rabbit antibodies?
Recombinant rabbit monoclonal antibodies have historically remained elusive due to intellectual property and technological barriers. Rabbit antibodies have unique advantages that make them ideal for research or diagnostic uses or as sources of therapeutic lead candidates. Due to higher diversity within antigen binding regions (CDRs), rabbit antibodies created from immunized sources have higher affinity compared to mouse antibodies, and rabbits can respond to antigens that are non-immunogenic in mice. Rabbit antibodies typically possess picomolar KD binding affinities, making affinity maturation unnecessary.
How are rabbit antibodies typically made?
Rabbit monoclonal antibodies are typically derived from hybridomas, which are commonly poorly characterized and can exhibit high batch-to-batch variability. Hybridoma library screening is limited in size (104-105), often missing rare clones that recognize important, less immunodominant epitopes. B-cell cloning methods suffer from similar limitations in library size and can lose additional clones due to poor gene recovery. Abwiz Bio’s proprietary recombinant rabbit monoclonal antibody development platform, RabWizTM combines the power of the rabbit immune response with the large library sizes (109-1011) of phage libraries.
But I thought rabbit Fabs do not perform well in phage display format?
Rabbit Fabs have a unique disulfide architecture that can cause expression problems in E. coli. Historically, this lead to the development of sub-optimal systems that do not use the full rabbit immune repertoire. This review article from 2017 recommends using chimeric rabbit/human Fab format-based phage display to avoid the expression issue. However, by using a novel patented antibody library construction method and a proprietary phage vector, the RabWizTM platform has been used to develop functional antibodies for hundreds of targets, including peptide, protein and cell-based antigens, using a fully rabbit Fab-phage display platform. See some of the unique characteristics and advantages of RabWizTM below.
Unique Advantages of RabWiz
A proprietary phage display vector compatible with unmodified rabbit Fabs
- The unique disulfide architecture of rabbit Fabs is known to cause toxicity in E. coli, which degrades the quality of phage libraries. Others have used sub-optimal systems that restrict usage of the full rabbit immune repertoire. Our highly optimized Fab-phage display vector expresses rabbit Fabs without modification.
A primer set that covers maximal VH gene diversity
- Through iterative improvement after creating dozens of rabbit libraries, we have created a primer set that covers the highest reported rabbit VH FR1 sequence diversity to date by including many FR1 sequences that are not currently in publicly available databases. Only by amplifying all antibody genes can you ensure that the entire rabbit immune repertoire is isolated during library construction.
WizAmpTM cloning (US 9,890,414)
- In our patented WizAmp™ procedure (US 9,890,414), a single non-gene-specific primer is used to amplify antibody genes prior to digestion and cloning into the library vector. This enables uniform amplification of antibody genes, allowing creation of unbiased, high quality libraries.
Abwiz Bio is pleased to perform contract work and to engage in co-development partnerships with a variety of research institutions and pharmaceutical companies. Abwiz Bio offers a variety of services that leverage our world-class, proprietary technology platform. Our partnership structure is highly flexible, and we are excited and interested in pursuing partnerships with pharmaceutical companies, research institutes or individual academic researchers. Call or e-mail us today to see how we can help you achieve your goals.
Recombinant monoclonal antibody development contracts
Hey! We’re looking for the next antibody development partner to join us Abwiz!
To learn how we can help you, contact us today!
Phage display based antibody generation
RabWizTM employs Abwiz Bio’s proprietary Needle-in-a-Haystack™ phage display technology and patented antibody library cloning method WizAmp™. This combines conventional animal immunization protocols with the robustness of phage display. These together enable the generation of antibodies with affinities typically attained only through hybridoma development, but doing so with the speed and diversity of phage display technology. The process allows for efficient, high throughput identification of rare binders with distinctive characteristics required for a specific application.
High affinity rabbit antibodies can be created in less than 6 months from the date of rabbit immunization. Abwiz Bio’s monoclonal antibody development platform is coupled with our friendly business terms (no royalties and minimal upfront fee).
Project Development timeline
Step I – Rabbit Immunization
Ia – Antigen preparation
Ib – Rabbit immunization
Depends on target
~ 2 months
Step II – Library Construction
Step III – Phage Selection
IIIa – Panning/screening
Fab ELISA screening data deliverable
CDR sequencing data deliverable
Affinity measurement deliverable (optional)
IIIb – Epitope masking selection (optional)
Repeat panning to obtain new clones
Step IV – Recombinant Antibody Small-Scale Production
Recombinant Antibody Small-Scale Production
IgG antibody ELISA data deliverable
~1 mg antibody deliverable
Step V – Recombinant Antibody Large-Scale Production
Recombinant Antibody Large-Scale Production
>3 mg antibody deliverable
Total time to create a new high-affinity recombinant rabbit antibody
Rabbit recombinant monoclonal antibodies represent a novel source of untapped potential and make great tools for use as research reagents or for high sensitivity immuno-detection in diagnostic assays. Rabbit antibodies are emerging as a unique source of lead candidates for therapeutic applications. Abwiz Bio has partnered with experts in rabbit antibody humanization, and our recombinant monoclonal antibody development platform is ideal for engineering humanized variants after the initial lead identification.
Is Abwiz Bio a Good fit for me?
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