Characterization of BK virus-specific antibodies in human sera by Western immunoblotting: use of a zwitterionic detergent for restoring the antibody-binding capacity of electroblotted proteins.
After initial problems related to denaturation of antigenic epitopes, we developed a Western immunoblotting method for the characterization of antibodies reacting with BK virus (BKV) structural polypeptides. When a zwitterionic detergent, Empigen BB, was added to the running buffer during electroblotting, the antibody-binding capacity of electrophoretically separated BKV polypeptides was partially restored. Antibodies reacting with different BKV antigens were detected and visualized by biotinylated anti-species-specific antibodies, peroxidase-conjugated streptavidine, and diaminobenzidine staining. Human sera containing anti-BKV antibodies reacted with VP1, but a serum containing antinuclear antibodies also reacted with VP4, -5 and -6 (histones). Serum from a rabbit inoculated with purified BKV reacted with VP1, and also with VP4, indicating that BKV inoculation may imply production of antibodies against histones.
Christie
K E
KE
Department of Microbiology, University of Tromsø, Norway.
Flaegstad
T
T
Traavik
T
T
D000914:Antibodies, Viral / Q000302:isolation & purification*; D001739:BK Virus / Q000276:immunology*; D003101:Collodion; D003902:Detergents; D004586:Electrophoresis; D006801:Humans; D009930:Organic Chemicals; D011120:Polyomavirus / Q000276:immunology*; D011485:Protein Binding
D000914:Antibodies, Viral; D003902:Detergents; D009930:Organic Chemicals; D003101:Collodion; C041772:empigen BB