Use of a zwitterionic detergent for the restoration of the antibody-binding capacity of electroblotted meningococcal outer membrane proteins.
A method is described for the partial restoration of the antibody-binding capacity of meningococcal class 2 outer membrane proteins (40-42 K molecular weight) following denaturation (dissociation) in boiling sodium dodecyl sulfate (SDS). The method relies on the presence of 0.1-0.4% zwitterionic detergent in the electrode buffer during the electroblot transfer of proteins from SDS-polyacrylamide gels to nitrocellulose paper. Meningococcal class 2 proteins which had lost their earlier capacity to bind mouse monoclonal antibodies in the normal blot procedure after SDS-polyacrylamide gel electrophoresis, bound monoclonal antibodies following the addition of the zwitterionic detergent to the blot buffer. Human post-vaccination anti-class 2 protein antibodies reacted with serotype 2a class 2 protein in a similar manner. This simple modification to the electroblot procedure proved helpful in identifying mouse monoclonal antibodies and human antibodies specific for native meningococcal class 2 proteins.
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins; D001666:Binding Sites, Antibody; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology*; D008581:Meningitis / Q000175:diagnosis / Q000276:immunology; D051379:Mice; D009345:Neisseria meningitidis / Q000276:immunology; D000644:Quaternary Ammonium Compounds; D011863:Radioimmunoassay; D012967:Sodium Dodecyl Sulfate
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D008565:Membrane Proteins; D000644:Quaternary Ammonium Compounds; C034730:zwittergent 3-12; D012967:Sodium Dodecyl Sulfate; D003101:Collodion