Integrating SpyCatcher/SpyTag covalent fusion technology into phage display workflows for rapid antibody discovery.

ABSTRACT: An early bottleneck in the rapid isolation of new antibody fragment binders using in vitro library approaches is the inertia encountered in acquiring and preparing soluble antigen fragments. In this report, we describe a simple, yet powerful strategy that exploits the properties of the SpyCatcher/SpyTag (SpyC/SpyT) covalent interaction to improve substantially the speed and efficiency in obtaining functional antibody clones of interest. We demonstrate that SpyC has broad utility as a protein-fusion tag partner in a eukaryotic expression/secretion context, retaining its functionality and permitting the direct, selective capture and immobilization of soluble antigen fusions using solid phase media coated with a synthetic modified SpyT peptide reagent. In addition, we show that the expressed

SUBMITTER: Fierle JK 

PROVIDER: S-EPMC6731262 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature