Immobilized pH gradient isoelectric focusing and immunoblotting for investigations of anti-Borrelia burgdorferi IgG antibodies.
Anti-Borrelia burgdorferi immunoglobulin G (IgG) responses in cerebrospinal fluid, serum, and joint fluid from Lyme disease patients were investigated by immobilized pH gradient (IPG) isoelectric focusing (IEF) in pH 4-10 and pH 4-7 gels. After focusing, the anti-B.-burgdorferi antibodies were blotted by affinity-driven transfer to antigen-coated polyvinylidene difluoride membranes (immunoblot) and the IgG antibodies were immunoenzymatically stained. IPG-IEF gels gave an excellent resolution of IgG and the immunoblot proved advantageous for the detection of anti-B. burgdorferi IgG antibodies. These antibodies, as judged from the electromigration characteristics, were found to contain oligoclonal as well as polyclonal subpopulations. This latter group included IgG antibodies that were inadequately resolved when separated by conventional carrier ampholyte IEF.
Cruz
M
M
Department of Neurology, Karolinska Institute, Huddinge, Sweden.
Sidén
A
A
D000907:Antibodies, Bacterial / Q000032:analysis*; D015153:Blotting, Western; D015748:Borrelia burgdorferi Group / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000032:analysis*; D007525:Isoelectric Focusing / Q000379:methods; D008193:Lyme Disease / Q000097:blood / Q000134:cerebrospinal fluid / Q000276:immunology; D013194:Staining and Labeling; D013582:Synovial Fluid / Q000276:immunology
D000907:Antibodies, Bacterial; D007074:Immunoglobulin G