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Determination of anti‐ADAMTS‐13 autoantibody titers in ELISA: Influence of ADAMTS‐13 presentation and autoantibody detection
Author(s) -
Dekimpe Charlotte,
Roose Elien,
Kangro Kadri,
Bonnez Quintijn,
Vandenbulcke Aline,
Tellier Edwige,
Kaplanski Gilles,
Feys Hendrik B.,
Tersteeg Claudia,
Männik Andres,
De Meyer Simon F.,
Vanhoorelbeke Karen
Publication year - 2021
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/jth.15297
Subject(s) - autoantibody , adamts , polyclonal antibodies , titer , antibody , monoclonal antibody , medicine , immunology , thrombospondin , metalloproteinase , matrix metalloproteinase
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.