PMID: 9168401May 1, 1997Paper

Human anti-Ro autoantibodies bind multiple conformational epitopes of 60-kD Ro autoantigen

Journal of Clinical Immunology
S C HuangJ B Harley

Abstract

A gel filtration method was developed to estimate the number of conformational epitopes on the 60-kD Ro antigen. Anti-Ro Fab or Fab' was incubated with native Ro antigen at different ratios and the Stokes radius molecular weight of complexes was estimated by gel filtration. Binding was saturated at 9 to 11 Fab molecules per bovine Ro molecule. Two additional Fab or Fab' were bound if human Ro was used as the antigen. Isolated Ro antigen/anti-Ro Fab complexes were evaluated for the relative proportion of antigen to antibody at saturation of antigen with antibody and thus stoichiometry was determined. This provided data supporting there being between 7 and 11 binding sites, results similar to those with the gel filtration method. Experiments carried out with anti-Ro monoclonal antibodies showed one binding site per molecule of 60-kD Ro. Therefore, we have developed methods to count conformational epitopes on autoantigens and have applied it to the Ro/anti-Ro system. The data indicate that multiple conformational epitopes can be bound simultaneously by polyclonal anti-Ro sera from patients with systemic lupus erythematosus.

Citations

Mar 10, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·M M Souto-CarneiroV Krenn
Nov 5, 2014·Antioxidants & Redox Signaling·Giuseppina BarreraFabrizio Gentile
Aug 31, 2002·Journal of Molecular Recognition : JMR·Carol S LutzJudith A James
Oct 31, 1998·Immunological Reviews·J A James, J B Harley
Apr 18, 2003·Clinical and Experimental Immunology·A W PurcellT P Gordon

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