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Immobilized pH gradient isoelectric focusing and immunoblotting for investigations of anti‐ Borrelia burgdorferi IgG antibodies
Author(s) -
Cruz Mabel,
Sidén Åke
Publication year - 1992
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150130147
Subject(s) - borrelia burgdorferi , isoelectric focusing , antibody , polyclonal antibodies , chemistry , microbiology and biotechnology , immunoglobulin g , antigen , borrelia , chromatography , biology , biochemistry , immunology , enzyme
Abstract 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.