By-passing in vitro screening--next generation sequencing technologies applied to antibody display and in silico candidate selection.

ABSTRACT: In recent years, unprecedented DNA sequencing capacity provided by next generation sequencing (NGS) has revolutionized genomic research. Combining the Illumina sequencing platform and a scFv library designed to confine diversity to both CDR3, >1.9 × 10(7) sequences have been generated. This approach allowed for in depth analysis of the library's diversity, provided sequence information on virtually all scFv during selection for binding to two targets and a global view of these enrichment processes. Using the most frequent heavy chain CDR3 sequences, primers were designed to rescue scFv from the third selection round. Identification, based on sequence frequency, retrieved the most potent scFv and valuable candidates that were missed using classical in vitro screening. Thus, by combining NGS

SUBMITTER: Ravn U 

PROVIDER: S-EPMC2995085 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature