Mammalian cell display for antibody engineering.
Antibody engineering has generally been carried out by displaying mouse or human antibodies or antibody fragments on the surface of microorganisms (phage, bacteria, and yeast). We have shown that mammalian cells can be used to display single-chain antibody fragments (scFvs) for affinity maturation. Using mammalian cell display one can isolate and engineer scFvs, Fabs, or whole IgGs for increased affinity and other specific biological functions. Here, we describe a mammalian cell display strategy to isolate high-affinity scFvs specific for CD22. Our strategy uses flow cytometry and human embryonic kidney 293T (HEK-293T) cells that are widely used for transient protein expression. Flow cytometry enhances the screen's sensitivity thereby allowing us to isolate high-affinity antibodies.
Ho
Mitchell
M
National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Pastan
Ira
I
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology; D007700:Kinetics; D018613:Microscopy, Confocal; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D017186:Titrimetry; D014162:Transfection; D014170:Transformation, Genetic
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D019151:Peptide Library