Genotyped functional screening of soluble Fab clones enables in-depth analysis of mutation effects.

ABSTRACT: Monoclonal antibodies (mAbs) and their fragments are widely used in therapeutics, diagnostics and basic research. Although display methods such as phage display offer high-throughput, affinities of individual antibodies need to be accurately measured in soluble format. We have developed a screening platform capable of providing genotyped functional data from a total of 9216 soluble, individual antigen binding fragment (Fab) clones by employing next-generation sequencing (NGS) with hierarchical indexing. Full-length, paired variable domain sequences (VL-VH) are linked to functional screening data, enabling in-depth analysis of mutation effects. The platform was applied to four phage display-selected scFv/Fab screening projects and one site-saturation VH affinity maturation project. Genotype

SUBMITTER: Oksanen S 

PROVIDER: S-EPMC10421887 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature