Affinity purification of antibodies from sera using polyvinylidenedifluoride (PVDF) membranes as coupling matrices for antigens presented by autoantibodies to triosephosphate isomerase.
Polyvinylidenedifluoride (PVDF) membranes covered with antigen can be used for affinity purification of antibodies from sera. These antibodies serve as an appropriate substitute for monoclonal antibodies when performing cytological and analytical experiments with the respective antigen. The usefulness of this method is demonstrated with the purification of IgM autoantibodies produced against human triosephosphate isomerase in acute Epstein-Barr virus infection. Compared with the original serum, the specific activity of the purified antibodies was up to 1400-fold higher, and the recovery of the anti-triosephosphate isomerase activity between 25 and 33%.
Ritter
K
K
Department of Medical Microbiology, Georg-August-University of Göttingen, F.R.G.
D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008567:Membranes, Artificial; D011145:Polyvinyls; D014305:Triose-Phosphate Isomerase / Q000276:immunology*; D014412:Tumor Virus Infections / Q000276:immunology; D014709:Vero Cells; D005465:Fluorocarbon Polymers
D001323:Autoantibodies; D007075:Immunoglobulin M; D008567:Membranes, Artificial; D011145:Polyvinyls; D014305:Triose-Phosphate Isomerase; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers