Purified recombinant 60 kD SS-A/Ro protein--an antigenic source for ELISA screening of patients with rheumatic diseases.
A 60 kD SS-A/Ro recombinant protein was expressed in E. coli cells and purified by the centrifugation procedure. Since the substrate contained a substantial amount of bacterial proteins it was not sufficient for ELISA. Therefore the material obtained by centrifugation was resolved on SDS-gel and the 60 kD region was excised and electro-eluted. Two hundred and nine sera from normal donors and patients with rheumatic diseases were analyzed by ELISA using the purified recombinant 60 kD SS-A/Ro protein. A comparison with Ouchterlony and Western blot revealed that ELISA proved to be a sensitive and specific method, which could discriminate between anti 60 kD antibodies and other autoantibodies. The present study showed that recombinant 60 kD may provide a purified source of antigen for ELISA while this method may serve as a useful tool for screening and evaluation of anti 60 kD antibodies and their relevance to clinical manifestations.
Ben-Chetrit
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W. M. Keck Autoimmune Disease Center, Scripps Clinic and Research Foundation, La Jolla, California.
Pollard
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KM
D001324:Autoantigens / Q000235:genetics / Q000302:isolation & purification*; D002498:Centrifugation; D004247:DNA / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D020733:RNA, Small Cytoplasmic; D011994:Recombinant Proteins / Q000302:isolation & purification; D012216:Rheumatic Diseases / Q000097:blood / Q000276:immunology*; D012261:Ribonucleoproteins; D000099207:SS-A Antigen
D001324:Autoantigens; D004247:DNA; D020733:RNA, Small Cytoplasmic; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human