Characterization of the antigen reactive with anti-Scl-70 antibodies and its application in an enzyme-linked immunosorbent assay.
The characteristics of the Scl-70 antigen (topoisomerase I) have been analyzed by means of autoantibodies. This antigen is a DNA-binding protein, dissociable from DNA at 0.3M NaCl and bound to a fraction of DNA that is very sensitive to nucleases. The molecular weight of the antigen is 105,000 daltons, whether dissociation conditions are used or not. Using chicken erythrocytes, and taking advantage of the strong interaction of the antigen with hydroxyapatite, we have designed a simple and fast purification protocol that allows the determination of anti-topoisomerase I antibodies by enzyme-linked immunosorbent assay.
Juarez
C
C
Department of Immunology, Hospital de la Santa Creu I Sant Pau, Barcelona, Spain.
Vila
J L
JL
Gelpi
C
C
Agusti
M
M
Amengual
M J
MJ
Martinez
M A
MA
Rodriguez
J L
JL
D000974:Antibodies, Antinuclear / Q000276:immunology* / Q000502:physiology; D001323:Autoantibodies / Q000276:immunology*; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D012595:Scleroderma, Systemic / Q000276:immunology*
D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D004264:DNA Topoisomerases, Type I