In vitro evidence that immunuaffinity-purified MOG contains immunogenic quantities of contaminating mouse IgG; techniques for producing Ig-free MOG.
Myelin oligodendrocyte glycoprotein (MOG) has attracted considerable attention due to its possible role in multiple sclerosis (MS). Previous studies have shown that MOG-reactive T cells are more commonly detected in MS patients than controls. In this report, we studied human MOG-reactive T cell clones generated from healthy individuals as well as MS patients. Our results indicate that immunoaffinity-purified MOG, which was routinely used in prior studies, is contaminated by anti-MOG antibody (mouse IgG), despite the inability to detect IgG by Western blotting. Here, we used continuous SDS-PAGE, which enabled us to isolate highly purified MOG devoid of contaminating mouse IgG.
Ohashi
T
T
Center for Neurologic Diseases, Laboratory of Molecular Immunology, Brigham and Women's Hospital and Harvard Medical School, 77 Avenue Louis Pasteur, HIM 786, Boston, MA 02115, USA.
Yukitake
M
M
Slavin
A
A
Krieger
J
J
Hafler
D A
DA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D002460:Cell Line; D002999:Clone Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D016130:Immunophenotyping; D007163:Immunosorbent Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D009103:Multiple Sclerosis / Q000276:immunology*; D009185:Myelin Proteins; D009186:Myelin Sheath / Q000737:chemistry; D018992:Myelin-Associated Glycoprotein / Q000276:immunology* / Q000302:isolation & purification* / Q000494:pharmacology; D063308:Myelin-Oligodendrocyte Glycoprotein; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D016207:Cytokines; D007074:Immunoglobulin G; C571326:MOG protein, human; C571327:Mog protein, mouse; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein