The orientation of β2GPI on the plate is important for the binding of anti-β2GPI autoantibodies by ELISA

G. Michael Iverson, Eiji Matsuura, Edward J. Victoria, Keith A. Cockerill, Matthew D. Linnik

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

(β2-glycoprotein I (β2GPI) is a plasma protein that plays an important role in the antigenic specificity of antiphospholipid autoantibodies (aPL). These antibodies are associated with an increased risk for thrombosis and recurrent foetal loss in humans. Crystallographic analysis of β2GPI showed that its five complement control protein (CCP) or 'sushi' domains are arranged in an elongated, fish-hook shape; yet the domain-specific location of epitopes recognized by these autoantibodies has remained the subject of considerable controversy. Investigators have used different forms of recombinant β2GPI and different ELISA methods to obtain conflicting results. One group mapped autoimmune epitopes to domain I using deletion mutants of β2GPI in a competitive inhibition ELISA on NUNC Maxisorp™ microplates. Another group mapped epitopes to domain IV using β2GPI with mutations in domain IV in a direct binding ELISA on polyoxygenated microplates. In an effort to resolve these discrepancies, a collaboration between the groups compared wildtype β2GPI with domain IV mutants in both types of ELISA. Autoantibodies bound very poorly to domain IV mutants coated on polyoxygenated plates, yet they bound very well to the same mutants coated on NUNC Maxisorp™ plates. The amount of protein adsorbed on to both types of plates was similar. In the competitive inhibition ELISA, no difference could be detected between wildtype β2GPI and domain IV mutants. These results strongly suggest that the orientation of β2GPI on the microplate, and not necessarily the lateral density, plays the predominant role in the binding of autoantibodies.

Original languageEnglish
Pages (from-to)289-297
Number of pages9
JournalJournal of Autoimmunity
Volume18
Issue number4
DOIs
Publication statusPublished - Jun 2002

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Autoantibodies
Glycoproteins
Enzyme-Linked Immunosorbent Assay
Epitopes
Blood Proteins
Complement System Proteins
Fishes
Thrombosis
Research Personnel
Mutation
Antibodies
Proteins

Keywords

  • Anti-β2GPI
  • Antiphospholipid syndrome

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

The orientation of β2GPI on the plate is important for the binding of anti-β2GPI autoantibodies by ELISA. / Iverson, G. Michael; Matsuura, Eiji; Victoria, Edward J.; Cockerill, Keith A.; Linnik, Matthew D.

In: Journal of Autoimmunity, Vol. 18, No. 4, 06.2002, p. 289-297.

Research output: Contribution to journalArticle

Iverson, G. Michael ; Matsuura, Eiji ; Victoria, Edward J. ; Cockerill, Keith A. ; Linnik, Matthew D. / The orientation of β2GPI on the plate is important for the binding of anti-β2GPI autoantibodies by ELISA. In: Journal of Autoimmunity. 2002 ; Vol. 18, No. 4. pp. 289-297.
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