A monoclonal antibody recognizing Golgi apparatus produced using affinity purified material from a patient with connective tissue disease.
Serum antibodies recognizing the Golgi apparatus have been reported in patients with connective tissue diseases, but little is known of their significance. Serum from a systemic lupus erythematosus patient with polymyositis was found to have high titers of anti-Golgi apparatus antibody. This serum recognized a 64 kD polypeptide in immunoblotting with HEp-2 cells. To verify that the 64 kD polypeptide was associated with the Golgi apparatus and to characterize which Golgi component was recognized, a monoclonal antibody was produced. IgG, isolated from this serum, was used in affinity chromatography to produce purified material which was used to generate a mouse monoclonal antibody. The monoclonal antibody had an indirect immunofluorescent pattern identical to that produced by the patient's serum, and similarly recognized a 64kD polypeptide in immunoblotting. A 59 kD polypeptide was also recognized by the monoclonal antibody, suggesting that the antigens recognized by the monoclonal and serum antibodies may be only partially identical. The antigen appears to be a glycoprotein and an integral component of the Golgi cisternae membranes.
Rossie
K M
KM
School of Dental Medicine, University of Pittsburgh, Pennsylvania.
Piesco
N P
NP
Charley
M R
MR
Oddis
C V
CV
Steen
V D
VD
Fratto
J
J
Deng
J S
JS
D000368:Aged; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006056:Golgi Apparatus / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology* / Q000473:pathology; D016253:Microscopy, Immunoelectron; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000276:immunology / Q000648:ultrastructure; D014909:Wheat Germ Agglutinins / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D014909:Wheat Germ Agglutinins