Cloning, Expression, and Purification of Monoclonal Antibodies in scFv-Fc Format
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.
Key wordsMonoclonal antibodies scFv-Fc format Hemagglutinin tag FreeStyle 293F cells Protein A affinity chromatography
This work was supported by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health. We thank Marko Modric for comments.