Display and selection of scFv antibodies on HEK-293T cells.
We describe a human cell display strategy to isolate high-affinity single-chain antibody fragments (scFvs) specific for CD22 for the treatment of B-cell malignancies. 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 scFvs. Using human cell display, one could isolate and engineer scFvs, single domains, Fabs, or whole IgGs for increased affinity and other biological functions.
Ho
Mitchell
M
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892-4264, USA.
Pastan
Ira
I
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology* / Q000302:isolation & purification*; D007668:Kidney / Q000196:embryology; D019151:Peptide Library; D051918:Sialic Acid Binding Ig-like Lectin 2 / Q000276:immunology*
D000911:Antibodies, Monoclonal; C497867:CD22 protein, human; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D051918:Sialic Acid Binding Ig-like Lectin 2