A new simple blot transfer method following cellogel electrophoresis which preserves antibody activity: application to the analysis of monoclonal and oligoclonal fluids.
Electrophoresis on gelatinized cellulose acetate (cellogel), followed by a simple blot transfer onto a nitrocellulose membrane, allows analysis of the electrophoretic pattern of specific antibody-active molecules by their binding of radiolabelled antigen. Using this non-denaturing method, it is easy to establish the monoclonal or oligoclonal nature of an antibody activity present in a serum or to control the cloning of a hybridoma cell line. Comparison between protein staining and autoradiography can afford a rough estimation of the relative affinities of different monoclonal antibodies towards the same antigen (in this case, the hepatitis B surface antigen). In addition, inhibition studies using other monoclonal antibodies directed against the same antigen may help to delineate the epitope specificity of antibodies.
J P
JP
Bouvet
J
J
Pillot
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000936:Antigen-Antibody Complex; D001345:Autoradiography; D002460:Cell Line; D004588:Electrophoresis, Cellulose Acetate / Q000379:methods; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D007118:Immunoassay; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D051379:Mice; D013194:Staining and Labeling
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006514:Hepatitis B Surface Antigens; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes