title: Monoclonal antibodies and a heterobifunctional reagent: a novel approach to the vectorial labeling of selected membrane proteins.
abstract: Many applications exist and others are envisioned for the chemical coupling of macromolecules to membrane proteins on the surface of mammalian cells. The ability to use antibody as a means to label and subsequently to follow the distribution of cell surface proteins is reported here. A new procedure is outlined for covalently coupling monoclonal antibodies to thiol-containing membrane proteins. The key reagent in the coupling reaction is the commercially available heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). The coupling proceeds in a simple two-step reaction in aqueous medium under very mild conditions. This results in a very efficient and stable attachment of anti-hapten antibodies to a selected set of cell surface proteins without any loss in cell viability and without denaturing the antibody molecule. The hapten-binding activity of the antibody is exploited to monitor the re-distribution of the antibody-labeled cell surface proteins periodically after the coupling reaction. The hapten binding activity can also be utilized to isolate membrane macromolecules via affinity chromatography.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D003432:Cross-Linking Reagents; D004912:Erythrocytes / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000378:metabolism; D011817:Rabbits; D012397:Rosette Formation; D012756:Sheep; D013388:Succinimides
chemical_list: D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate