Autoantibodies against topoisomerase I detected with the natural enzyme and overlapping recombinant peptides.
Antibodies against topoisomerase I (anti-topo I, anti-Scl-70) are regarded as a marker of systemic sclerosis. The various frequencies of anti-topo I detected in those patients depends at least in part on the test design and the kind of the antigen used. We therefore analyzed three overlapping recombinant topo I fragments (N-terminal, center and C-terminal part of the molecule) covering the full length of the enzyme for substitution of highly purified natural antigens (n-topo I) in ELISA for antibody screening. 49 of 50 sera reacting with n-topo I in ELISA also recognized the recombinant C-terminal topo I fragment under identical test conditions, 37 sera recognized the recombinant center and two sera the recombinant N-terminal peptide. All sera reactive with the N-terminal and center peptide reacted with the recombinant C-terminus which therefore may substitute for the natural antigen. In immunoblot assays 92% (46/50) of the sera reacted with n-topo I and 86% (43/50) with the recombinant C-terminal peptide. Immunoblots therefore seem to be less sensitive for detecting anti-topo I antibodies than ELISA regardless the source of the antigen used. In a screening of 696 sera submitted for routine antibody tests the recombinant peptide ELISA on the other hand detected two sera which did not react with n-topo I in ELISA. Because of the high rate of agreement within the results obtained with the two antigens, n-topo I can be substituted by the recombinant peptide ELISA allowing better standardization and interlaboratory comparison of results.
Seelig
H P
HP
Institute of Immunology and Molecular Genetics, Karlsruhe, Germany.
Schröter
H
H
Ehrfeld
H
H
Renz
M
M
D000818:Animals; D000941:Antigens / Q000032:analysis; D001323:Autoantibodies / Q000032:analysis*; D002417:Cattle; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000276:immunology*; D018076:DNA, Complementary / Q000097:blood / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D010446:Peptide Fragments / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000276:immunology; D012595:Scleroderma, Systemic / Q000097:blood / Q000276:immunology; D013950:Thymus Gland / Q000201:enzymology
D000941:Antigens; D001323:Autoantibodies; D018076:DNA, Complementary; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004264:DNA Topoisomerases, Type I