A Shorter Route to Antibody Binders via Quantitative in vitro Bead-Display Screening and Consensus Analysis.

ABSTRACT: Affinity panning of large libraries is a powerful tool to identify protein binders. However, panning rounds are followed by the tedious re-screening of the clones obtained to evaluate binders precisely. In a first application of Bead Surface Display (BeSD) we show successful in vitro affinity selections based on flow cytometric analysis that allows fine quantitative discrimination between binders. Subsequent consensus analysis of the resulting sequences enables identification of clones that bind tighter than those arising directly from the experimental selection output. This is demonstrated by evolution of an anti-Fas receptor single-chain variable fragment (scFv) that was improved 98-fold vs the parental clone. Four rounds of quantitative screening by fluorescence-activated cell sorting o

SUBMITTER: Mankowska SA 

PROVIDER: S-EPMC5098251 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature