Determination of anti-ADAMTS-13 autoantibody titers in ELISA: Influence of ADAMTS-13 presentation and autoantibody detection

Charlotte Dekimpe; Elien Roose; Kadri Kangro; Quintijn Bonnez; Aline Vandenbulcke; Edwige Tellier; Gilles Kaplanski; Hendrik B Feys; Claudia Tersteeg; Andres Männik; Simon F De Meyer; Karen Vanhoorelbeke

doi:10.1111/jth.15297

Determination of anti-ADAMTS-13 autoantibody titers in ELISA: Influence of ADAMTS-13 presentation and autoantibody detection

J Thromb Haemost. 2021 Sep;19(9):2248-2255. doi: 10.1111/jth.15297. Epub 2021 Jul 14.

Authors

Affiliations

¹ Laboratory for Thrombosis Research, IRF Life Sciences, KU Leuven Campus Kulak Kortrijk, Kortrijk, Belgium.
² Icosagen Cell Factory OÜ, Kambja vald, Tartumaa, Estonia.
³ C2VN, INSERM, INRAE, Aix Marseille Universite, Marseille, France.
⁴ Hôpital de la Conception, Service de médecine interne, APHM, C2VN, INSERM, INRAE, Aix Marseille Universite, Marseille, France.
⁵ Transfusion Research Center, Belgian Red Cross-Flanders, Ghent, Belgium.
⁶ Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium.

PMID: 33728786
DOI: 10.1111/jth.15297

Abstract

Background: Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is caused by inhibitory and/or clearing anti-ADAMTS-13 (A Disintegrin and Metalloprotease with ThromboSpondin type 1 repeats, member 13) autoantibodies. To determine the presence and total level of anti-ADAMTS-13 autoantibodies, commercial and in-house developed ELISAs are performed. However, different ELISA methods vary in relation to the presentation of recombinant (r)ADAMTS-13 and the detection method of the anti-ADAMTS-13 autoantibodies. Currently, the influence of those different approaches on anti-ADAMTS-13 autoantibody titers is not known.

Objectives: To assess the influence of different ADAMTS-13 presentation- and autoantibody detection methods on anti-ADAMTS-13 autoantibody titers in ELISA.

Materials/methods: Anti-ADAMTS-13 autoantibody titers from 18 iTTP patients were determined using four different set-ups of anti-ADAMTS-13 autoantibody ELISAs. The ELISAs varied in the used presentation of rADAMTS-13 (directly coated full-length rADAMTS-13, directly coated rMDTCS and rT2C2, or antibody-captured full-length rADAMTS-13) and the detection antibodies (polyclonal anti-human IgG or monoclonal anti-human IgG_1-4 antibodies).

Results: Strong correlations between the different anti-ADAMTS-13 autoantibody ELISA approaches were observed, when using polyclonal anti-human IgG detection antibodies recognizing all IgG subclasses similarly, independent of the method of rADAMTS-13 presentation. Anti-ADAMTS-13 autoantibody titers correlated less when using a mixture of monoclonal anti-human IgG_1-4 , because not all IgG subclasses were recognized with similar affinities.

Conclusion: Anti-ADAMTS-13 autoantibody levels using different methods of rADAMTS-13 presentation strongly correlate. However, the levels of anti-ADAMTS-13 autoantibodies are highly dependent on the detection antibody used, which should detect all IgG subclasses (IgG_1-4 ) equally well.

Keywords: ADAMTS-13 protein; antibodies; diagnosis; thrombotic microangiopathies; thrombotic thrombocytopenic purpura.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADAMTS13 Protein
Autoantibodies
Enzyme-Linked Immunosorbent Assay
Humans
Purpura, Thrombotic Thrombocytopenic* / diagnosis
Thrombospondin 1*

Substances

Autoantibodies
Thrombospondin 1
ADAMTS13 Protein