We have developed a novel method using NGS and bioinformatics to identify the full antigen-specific repertoire of heavy-chain antibodies generated by UniRat after immunization. We have also developed unique gene assembly methods to convert the antibody repertoire sequence information into large collections of fully-human heavy-chain antibodies that can be screened for a variety of functions.
We believe that success in therapeutic antibody discovery and development is greatly enabled by starting with a large and comprehensive set of sequence-defined antibody candidates.
Example 1: NGS analysis reveals expanded VH sequence lineages
The degree of red (high) or blue (low) in the heatmap indicates expanded VH sequence families due to immunization. Some highly ranked VH sequences are unique to a single animal. Other highly ranked VH sequences appear in more than one animal suggesting convergent selection of those sequence familes.
Example 2: Primary screen in UniAb format identifies many antigen-specific VH sequence families
A total of 327 high-frequency VH sequences were selected for recombinant expression and screening. UniAbs were screened by ELISA for protein binding and flow cytometry for cell binding.
327 unique VH sequences, from >250 CDR3 sequence families
31% (100) human protein ELISA positive