This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Walton, S.P. Articles by Harris, E.N. Search for Related Content PubMed PubMed Citation Articles by Walton, S.P. Articles by Harris, E.N. Social Bookmarking                         What's this?

Demonstration of antiphospholipid antibody heterogeneity by phospholipid column chromatography and salt gradient elution techniques

Department of Medicine, Division of Rheumatology, University of Louisville, Louisville, KY 40292

S.S. Pierangeli

Antiphospholipid Standardization Laboratory, University of Louisville, Louisville, KY 40292

A. Campbell

Antiphospholipid Standardization Laboratory, University of Louisville, Louisville, KY 40292

E. Klein

Department of Medicine, Kidney Disease Program, University of Louisville, Louisville, KY 40292

B. Burchitt

Antiphospholipid Standardization Laboratory, University of Louisville, Louisville, KY 40292

E.N. Harris

Department of Medicine, Division of Rheumatology, University of Louisville, Louisville, KY 40292

Antiphospholipid (aPL) antibodies are associated with thrombosis, recurrent abortions and thrombocytopenia. Studies to determine the mechanisms of action of these antibodies have been hindered by their heterogeneity and limited availability of techniques to isolate and characterize subgroups of the antibodies. We report a new phospholipid affinity chromatography method which enables separation of antiphospholipid positive sera into more than one antibody subpopulation. Sera from five patients with complications of the antiphospholipid syndrome (APS) were studied. Each serum was applied to chromatography columns prepared by coating polystyrene beads (diameter 100 Å) with phosphatidylserine (PS) or cardiolipin (CL). A linear salt gradient (0.03-1.0 M NaCl) was used for elution. Eluates were analyzed for phospholipid binding and for inhibition of the prothrombin-thrombin conversion reaction. Each sample yielded two to three peaks for CL and PS affinity columns. Molarities at which peaks were eluted differed between samples. For individual samples, molarities at which peaks were eluted differed between CL and PS columns. These data suggest that aPL antibodies are heterogenous, with differences existing between patients and even within single serum samples. Subpopulations differed in their avidities for CL and PS but generally all had prothrombinase inhibitory activity.

Key Words: anticardiolipin antibodies • antiphospholipid antibodies • lupus anticoagulant • antiphospholipid syndrome • affinity chromatography

Lupus, Vol. 4, No. 4, 263-271 (1995)
DOI: 10.1177/096120339500400406


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?