Identification of the specificity of isolated phage display single-chain antibodies using yeast two-hybrid screens.
A method is described for the identification of the antigen recognised by an scFv isolated from an antibody phage display library using selection against a complex mixture of proteins (e.g. intact cells, purified cell surface membranes, and tissue sections). The method takes advantage of a yeast two-hybrid system that additionally allows for reorganization of post-translational modifications to the bait and target proteins. This technique is therefore especially useful for identifying surface-expressed antigens.
Rasmussen
Nicolaj
N
Institute of Medical Biology, University of Southern Denmark, Winsloewparken 25, 3, DK-5000, Odense, Denmark.
Ditzel
Henrik J
HJ
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000276:immunology; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology / Q000302:isolation & purification*; D019151:Peptide Library; D011499:Protein Processing, Post-Translational; D014169:Transformation, Bacterial; D020798:Two-Hybrid System Techniques
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007135:Immunoglobulin Variable Region; D019151:Peptide Library