Phage Display Antibody Library Development
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Phage Display Antibody Library Development & Other Technologies
Abwiz Bio was founded in 2012 to leverage the power of state-of-the-art recombinant antibody technologies to develop research tools and diagnostics and to co-develop therapeutics. The founding and executive team at Abwiz Bio has extensive experience in antibody engineering technologies, from discovery and lead identification through commercialization. Our company differentiates itself from others by our state of the art rabbit immunization system, and by our patented phage display antibody library development and panning/screening systems.
Rabbit monoclonal antibodies possess higher affinity (picomolar vs. nanomolar KD) and higher diversity in antigen-binding regions over their mouse counterparts, and can generate antibodies against antigens and epitopes that are non-immunogenic in mice, making them more useful in downstream applications. Traditionally, isolation of rabbit monoclonal antibodies is performed by hybridoma technology, a labor-intensive process that is strongly biased for selection of antibodies that recognize immunodominant epitopes. Hybridoma-derived antibodies can exhibit high batch-to-batch variability, leading a number of researchers to call for the use of only stringently characterized recombinant monoclonal antibody products. The hybridoma approach suffers from significant disadvantages, and intellectual property protection severely limits the use of rabbit hybridoma technology for companies looking to enter this market. Additionally, rabbit B cells fuse with cancer cells with very low efficiency, and the resulting hybridomas tend to have poor viability. Finally, hybridoma technology is limited to only screening a small proportion of the entire antibody repertoire and thus often misses rare clones recognizing important, less immunodominant epitopes.
Despite their many advantages including higher affinity and specificity, the production of recombinant rabbit monoclonal antibodies has been difficult to date due to a well-known Fab toxicity problem in E. coli. At Abwiz Bio, our patented WizAmpTM(US 9,890,414) amplification procedure creates rabbit phage display antibody libraries with larger diversity and higher quality compared to any other available methods. Our antibody engineering platform, RabwizTM, has proven effective at the selection and creation of recombinant rabbit antibodies from immunized libraries due to the use of a proprietary, highly optimized Fab/phage expression vector.
Abwiz Bio's Needle-in-a-HaystackTM technology combines a differentiating and proprietary antibody library cloning method, WizAmpTM(US 9,890,414), with conventional animal immunization protocols and 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. Our patented RabwizTM phage display antibody library development platform is based on (1) capturing the highest antibody diversity reported to date from rabbit immune libraries through our WizAmpTM (US 9,890,414) amplification method and (2) the use of a highly-optimized, proprietary Fab/phage vector that allows rabbit Fab expression in bacterial cells without antibody structure modification.
The process additionally allows for efficient, high throughput identification of rare binders with distinctive characteristics required for a specific application. Furthermore, the Needle-in-a-HaystackTM technology allows Abwiz Bio to generate antibodies to targets that have traditionally been difficult to attain, such as GPCRs. Abwiz Bio's phage display antibody library platform is coupled with our partner-friendly business terms (no royalties and minimal upfront fees), providing significant advantages to partners in seeking specific antibodies.
Proprietary phage display antibody library cloning method, WizAmp™ (US 9,890,414)
Our proprietary antibody cloning method (patent pending) enables the robust amplification of antibody genes without bias from the immune source. This greatly enhances the diversity of the resultant library and leads to the discovery of very rare and important antibodies.
1. Phage display enables the isolation of a panel of antibodies that recognize multiple epitopes
- Antibodies that recognize multiple epitopes on a small molecule
- Agonistic and antagonistic antibodies
- Neutralizing and non-neutralizing antibodies
Antibodies against an immuno-dominant epitope are not always superior functionally.
Epitope masking, which can be easily performed when using phage display technology, is very effective in the isolation of antibodies that recognize different epitopes on the same antigen.
2. Phage display enables the isolation of antibodies that specifically recognize highly homologous antigens
- Proteins with high homology in amino acid sequence
- Post translational modifications (e.g. phospho-, methylation, and glycoyslation-specific antibodies, etc.)
Rabbit monoclonal antibody development platform, Rabwiz™
Abwiz Bio's proprietary rabbit monoclonal antibody development platform (RabwizTM) allows for the rapid selection of specific and high-affinity binders from highly diverse (109 to 1011 in size) libraries. This technology has numerous advantages over traditional rabbit antibody engineering methods:
- Our patented WizAmpTM (US 9,890,414) phage display antibody library cloning method creates large, high quality libraries from immunized rabbits by unbiased amplification of antibody genes.
- Our proprietary phage display vector readily incorporates rabbit Fabs without additional modification. It is well known that rabbit Fabs are toxic in E. coli. This has resulted in the creation of non-optimal systems that do not utilize the full rabbit immune repertoire, including the expensive Basiela rabbit strain that lacks disulfide-containing kappa chains, construction of chimeric Fabs containing human constant domains or significant engineering in the scFv format. However, our highly optimized vector expresses rabbit Fabs at high levels without modification.
- We have developed a specialized software program that analyzes and groups amino acid sequences, allowing for rapid identification of lead candidates from hundreds of clones after activity-based screening. This assures that even a single rare but possibly important clone is not overlooked.
- We have streamlined cloning and expression of lead candidates in mammalian systems in various formats (e.g. Fab, F(ab)'2, and chimeric IgG, etc…) to facilitate downstream functional assays and further development into animal studies.
Abwiz Bio's phage display antibody library technology (RabwizTM) offers an improved alternative to mouse monoclonal or rabbit polyclonal antibodies. Abwiz Bio strives to become a leading provider of recombinant rabbit monoclonal antibodies for reagent use, diagnostic development and therapeutic use. We also offer antibody development services to our clients under a variety of funding options including fee-for-service or milestone payments.
(peptide, protein, cells, lipid, etc.)
Antigens are prepared for immunization depending on the nature of the antigens.
Immunization and serum test
Immunization in rabbits is performed using the antigens previously mentioned and the serum titers are tested by various assays. (e.g. ELISA, flow cytometry)
Libraries are constructed using our unique cloning method (WizAmpTM) with proprietary primer sets to cover maximum diversity from immune source.
Phage display panning
Specific clones are selected using our Needle-in-a-HaystackTM strategy.
Sequence analysis and direct cloning into IgG expression vectors
Amino acid sequences of positive clones are analyzed using our proprietary sequence analysis program and lead candidates are directly cloned into IgG expression vectors.