Cloning, expression, and purification of monoclonal antibodies in scFv-Fc format

Abstract

This protocol describes the generation of monoclonal antibodies in single-chain variable fragment (scFv)-Fc format. It includes the cloning of the scFv-Fc expression cassette into a mammalian expression vector followed by transient transfection of mammalian cells and purification by protein A affinity chromatography. The protocol is intended for applications in basic and preclinical research that require rapid access to milligram amounts of protein. © 2012 Springer Science+Business Media, LLC.