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.
Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Jiahui
J
Yang
Christoph
C
Rader
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D006801:Humans; D057127:Single-Chain Antibodies / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D057127:Single-Chain Antibodies