To bring revolutionary speed and diversity to antibody generation, Abzyme developed yeast-based technology platforms mimicking mammalian antibody diversification processes and applied the platform for developing human antibodies.
Advantages of Abzyme’s human Fab and IgG antibody discovery platform include:
• Unlimited diversity: Random combination of heavy chain and light chain libraries by yeast mating and inducible diversification by hypermutation.
• Eukaryotic-based in vitro discovery system: Filters for well expressing antibody and manufacturable candidates to difficult targets such as, non-immunogenic and toxic antigens as well as cell surface antigens for membrane protein antibody production.
• In vivo antigen-directed antibody maturation: Mimicking mammalian antibody diversification processes in combination with selection and FACS sorting
• Speed of development: Each discovery/optimization project finishes within weeks from target to fully human F(ab) or IgG antibodies
• FACS-based approach identifies relevant antibodies: Advanced process for identifying antibodies with desired attributes
• Advanced antibody optimization capabilities: Rapidly optimize leads into best-in-class therapeutic candidates
• Consistency across production batches: Recombinant monoclonal antibody with known amino acid sequences
A highly diversified naïve human Fab yeast display library is used that was formed by random combination of heavy and light chains by yeast mating of two sub-libraries each with diversity levels >10^7 and derived from more than 500 donors. Theoretically the diversity level of this Fab library is on the order of 10^14. The library is subjected to further diversification using Abzyme’s proprietary inducible hypermutation platform and selection process. After three diversification/enrichment cycles, yeast cells displaying high affinity human Fab to the target antigen are twice sorted by FACS to isolate single clones. Additional enrichment and FACS sorting may be applied to increase antibody affinity or to select clones with additional activities. FACS-sorted single yeast display Fab clone candidates are further validated by cytoflow analysis using various antigen concentrations to rank order clones for affinity to antigens. Confirmed clones will be switched to a secretion mode for antibody production or transferred into mammalian cell expression system for IgG production of final antibodies.