Direct immunoprecipitation of antigen with phage displaying immunoglobulin fragment.
Phage libraries may display hormones, receptors, antibody fragments, etc., by fusion to phage envelope proteins. This report describes the direct precipitation of phage-Fab-antigen complexes by polyethylene glycol precipitation, resulting in highly selective and efficient recovery of antigen from complex mixtures without nonspecific protein contamination. The method demonstrates efficiency and specific recovery of phage-Fab-antigen complexes from a background of a complex mixture of unrelated proteins as may occur in the analysis of biological specimens. This simple, fast, and effective method allows isolation and characterization of target antigens, with no need to further process Fab or sFv, and may reasonably be extended to isolate any interacting partner molecule for any displayed protein.
Telleman
P
P
Beth Israel Deaconess Medical Center, Boston, MA, USA.
Kingsbury
G A
GA
Junghans
R P
RP
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism; D000941:Antigens / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D003090:Coliphages / Q000276:immunology* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007128:Immunoglobulin Fragments / Q000378:metabolism*; D051379:Mice; D019151:Peptide Library; D011092:Polyethylene Glycols; D011233:Precipitin Tests / Q000379:methods*; D015375:Receptors, Interleukin-2 / Q000276:immunology; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D015375:Receptors, Interleukin-2; C055966:immunoglobulin Fv; D011092:Polyethylene Glycols