Use of a zwitterionic detergent for the restoration of the antibody-binding capacity of electroblotted meningococcal outer membrane proteins

J Immunol Methods. 1984 Feb 24;67(1):1-11. doi: 10.1016/0022-1759(84)90080-2.

Abstract

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.

MeSH terms

  • Animals
  • Antigens, Bacterial / analysis
  • Antigens, Bacterial / immunology*
  • Bacterial Outer Membrane Proteins
  • Binding Sites, Antibody*
  • Collodion
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Humans
  • Membrane Proteins / analysis
  • Membrane Proteins / immunology*
  • Meningitis / diagnosis
  • Meningitis / immunology
  • Mice
  • Neisseria meningitidis / immunology
  • Quaternary Ammonium Compounds*
  • Radioimmunoassay
  • Sodium Dodecyl Sulfate

Substances

  • Antigens, Bacterial
  • Bacterial Outer Membrane Proteins
  • Membrane Proteins
  • Quaternary Ammonium Compounds
  • zwittergent 3-12
  • Sodium Dodecyl Sulfate
  • Collodion