We intend to refine the affinity-reagent discovery process so that it is not only cheaper and faster than currently possible, but also so that we produce reagents that are more versatile and useful than current monoclonal antibodies. We propose to use in vitro combinatorial recombination of pre-defined synthetic complementarity determining regions (CDRs) of single chain variable fragment (scFv) antibodies to synthesize a novel type of highly diverse synthetic display library. We have termed this new library-type as a 'pre-defined CDR'(PDC) library. This novel library system will have properties that will also enable it to greatly facilitate both the downstream affinity-reagent selection and the maturation processes. Methods for testing this library using microfluidics and emulsion screening are discussed.
We intend to refine the affinity-reagent discovery process so that it is not only cheaper and faster than currently possible, but also so that we produce reagents that are more versatile and useful than current monoclonal antibodies. A novel library system is proposed that will have properties that will enable it to greatly facilitate both the downstream affinity-reagent selection and the maturation processes. Methods for testing this library using microfluidics and emulsion screening are discussed. This library and the method proposed recapitulates in vitro the way antibodies in vivo are enriched as a response to an antigen assault. But we will be able to achieve a comparable analysis on the much larger recombinant libraries (for example 109-1012) we can generate in vitro. Completion of this method has the potential to save time and money and result in a better product over current methods for recombinant affinity-reagent generation.
| Batonick, Melissa; Holland, Erika G; Busygina, Valeria et al. (2016) Platform for high-throughput antibody selection using synthetically-designed antibody libraries. N Biotechnol 33:565-73 |
