Anti‐ADAMTS13 autoantibodies in immune‐mediated thrombotic thrombocytopenic purpura do not hamper ELISA‐based quantification of ADAMTS13 antigen

Background The biological diagnosis of immune-mediated thrombotic thrombocytopenic purpura (iTTP) is based on determination of ADAMTS13 activity (

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Published in:Journal of thrombosis and haemostasis 2020-01, Vol.18 (4), p.985-990
Main Authors: Dekimpe, Charlotte, Roose, Elien, Tersteeg, Claudia, Joly, Bérangère, Dewaele, Aurelie, Horta, Sara, Pareyn, Inge, Vandenbulcke, Aline, Deckmyn, Hans, Feys, Hendrik, Tellier, Edwige, Kaplanski, Gilles, Scully, Marie, Coppo, Paul, de Meyer, Simon, Veyradier, Agnès, Vanhoorelbeke, Karen
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Cardiology and cardiovascular system
Human health and pathology
Life Sciences
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container_end_page 990
container_issue 4
container_start_page 985
container_title Journal of thrombosis and haemostasis
container_volume 18
creator Dekimpe, Charlotte
Roose, Elien
Tersteeg, Claudia
Joly, Bérangère
Dewaele, Aurelie
Horta, Sara
Pareyn, Inge
Vandenbulcke, Aline
Deckmyn, Hans
Feys, Hendrik
Tellier, Edwige
Kaplanski, Gilles
Scully, Marie
Coppo, Paul
de Meyer, Simon
Veyradier, Agnès
Vanhoorelbeke, Karen
description Background The biological diagnosis of immune-mediated thrombotic thrombocytopenic purpura (iTTP) is based on determination of ADAMTS13 activity (
doi_str_mv 10.1111/jth.14747
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ADAMTS13 antigen levels are not routinely measured in iTTP patients, but studies have shown that antigen levels are a valuable prognostic factor. Objectives To (a) report the validation of our in-house developed ADAMTS13 antigen enzyme-linked immunosorbent assay (ELISA) and determine ADAMTS13 antigen in a large cohort of healthy donor and iTTP patient plasma samples; and (b) to investigate whether ADAMTS13 antigen determination is not disturbed by the presence of anti-ADAMTS13 autoantibodies.Methods Our in-house ADAMTS13 antigen ELISA was validated in terms of sensitivity, repeatability, and reproducibility. ADAMTS13 antigen levels were determined in plasma samples from 423 healthy donors and 112 acute iTTP patients. Purified IgGs from iTTP patients were added to normal human plasma to determine whether anti-ADAMTS13 autoantibodies hampered ADAMTS13 antigen determination.Results Our in-house ADAMTS13 antigen ELISA has a detection limit of 3% and low intra-assay (coefficient of variation, %CV &lt; 10%) and inter-assay (%CV &lt; 18%) variability. ADAMTS13 antigen levels were significantly reduced (P &lt; .0001) in acute iTTP patients (15 +/- 18%) compared to healthy donors (101 +/- 18%). The anti-ADAMTS13 autoantibodies in plasma of iTTP patients did not impede ADAMTS13 antigen determinations using our in-house ELISAConclusions Our in-house ADAMT13 antigen ELISA is a powerful tool to correctly determine ADAMTS13 antigen levels in iTTP patients, which supports routine ADAMTS13 antigen measurements in these patients to have better insight into disease prognosis.</description><identifier>ISSN: 1538-7933</identifier><identifier>EISSN: 1538-7836</identifier><identifier>DOI: 10.1111/jth.14747</identifier><language>eng</language><publisher>Wiley</publisher><subject>Cardiology and cardiovascular system ; Human health and pathology ; Life Sciences</subject><ispartof>Journal of thrombosis and haemostasis, 2020-01, Vol.18 (4), p.985-990</ispartof><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0002-6380-6349 ; 0000-0003-2288-8277 ; 0000-0002-9078-9070 ; 0000-0003-3044-5388 ; 0000-0001-7773-5003 ; 0000-0002-9078-9070 ; 0000-0001-7773-5003 ; 0000-0003-2288-8277 ; 0000-0003-3044-5388 ; 0000-0002-6380-6349</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://hal.inrae.fr/hal-04017856$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Dekimpe, Charlotte</creatorcontrib><creatorcontrib>Roose, Elien</creatorcontrib><creatorcontrib>Tersteeg, Claudia</creatorcontrib><creatorcontrib>Joly, Bérangère</creatorcontrib><creatorcontrib>Dewaele, Aurelie</creatorcontrib><creatorcontrib>Horta, Sara</creatorcontrib><creatorcontrib>Pareyn, Inge</creatorcontrib><creatorcontrib>Vandenbulcke, Aline</creatorcontrib><creatorcontrib>Deckmyn, Hans</creatorcontrib><creatorcontrib>Feys, Hendrik</creatorcontrib><creatorcontrib>Tellier, Edwige</creatorcontrib><creatorcontrib>Kaplanski, Gilles</creatorcontrib><creatorcontrib>Scully, Marie</creatorcontrib><creatorcontrib>Coppo, Paul</creatorcontrib><creatorcontrib>de Meyer, Simon</creatorcontrib><creatorcontrib>Veyradier, Agnès</creatorcontrib><creatorcontrib>Vanhoorelbeke, Karen</creatorcontrib><title>Anti‐ADAMTS13 autoantibodies in immune‐mediated thrombotic thrombocytopenic purpura do not hamper ELISA‐based quantification of ADAMTS13 antigen</title><title>Journal of thrombosis and haemostasis</title><description>Background The biological diagnosis of immune-mediated thrombotic thrombocytopenic purpura (iTTP) is based on determination of ADAMTS13 activity (&lt;10%) and anti-ADAMTS13 autoantibodies. ADAMTS13 antigen levels are not routinely measured in iTTP patients, but studies have shown that antigen levels are a valuable prognostic factor. Objectives To (a) report the validation of our in-house developed ADAMTS13 antigen enzyme-linked immunosorbent assay (ELISA) and determine ADAMTS13 antigen in a large cohort of healthy donor and iTTP patient plasma samples; and (b) to investigate whether ADAMTS13 antigen determination is not disturbed by the presence of anti-ADAMTS13 autoantibodies.Methods Our in-house ADAMTS13 antigen ELISA was validated in terms of sensitivity, repeatability, and reproducibility. ADAMTS13 antigen levels were determined in plasma samples from 423 healthy donors and 112 acute iTTP patients. Purified IgGs from iTTP patients were added to normal human plasma to determine whether anti-ADAMTS13 autoantibodies hampered ADAMTS13 antigen determination.Results Our in-house ADAMTS13 antigen ELISA has a detection limit of 3% and low intra-assay (coefficient of variation, %CV &lt; 10%) and inter-assay (%CV &lt; 18%) variability. ADAMTS13 antigen levels were significantly reduced (P &lt; .0001) in acute iTTP patients (15 +/- 18%) compared to healthy donors (101 +/- 18%). The anti-ADAMTS13 autoantibodies in plasma of iTTP patients did not impede ADAMTS13 antigen determinations using our in-house ELISAConclusions Our in-house ADAMT13 antigen ELISA is a powerful tool to correctly determine ADAMTS13 antigen levels in iTTP patients, which supports routine ADAMTS13 antigen measurements in these patients to have better insight into disease prognosis.</description><subject>Cardiology and cardiovascular system</subject><subject>Human health and pathology</subject><subject>Life Sciences</subject><issn>1538-7933</issn><issn>1538-7836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNqVjU1OwzAQhS0EEuVnwQ1my6LFxkmTLi1aVKSyavfRJHHIVLUdHAepux6BFQfkJLgSFWtGI83TN2_mMXYn-ETEetiGdiKSLMnO2EikMh9nuZyen_RMykt21fdbzsUsfeQj9qVsoO_Dp5qr181aSMAhOIysdDXpHsgCGTNYHT1G14RB1xBa70zpAlUnWe2D67SNoBt8bITagXUBWjSd9rBYvaxVfFFiH-_fh2NCQxUGchZcA3_xcfGm7Q27aHDX69vfec3unxebp-W4xV3ReTLo94VDKpZqVRwZT7jI8nT6IeR_vD91bWQ1</recordid><startdate>20200113</startdate><enddate>20200113</enddate><creator>Dekimpe, Charlotte</creator><creator>Roose, Elien</creator><creator>Tersteeg, Claudia</creator><creator>Joly, Bérangère</creator><creator>Dewaele, Aurelie</creator><creator>Horta, Sara</creator><creator>Pareyn, Inge</creator><creator>Vandenbulcke, Aline</creator><creator>Deckmyn, Hans</creator><creator>Feys, Hendrik</creator><creator>Tellier, Edwige</creator><creator>Kaplanski, Gilles</creator><creator>Scully, Marie</creator><creator>Coppo, Paul</creator><creator>de Meyer, Simon</creator><creator>Veyradier, Agnès</creator><creator>Vanhoorelbeke, Karen</creator><general>Wiley</general><scope>1XC</scope><orcidid>https://orcid.org/0000-0002-6380-6349</orcidid><orcidid>https://orcid.org/0000-0003-2288-8277</orcidid><orcidid>https://orcid.org/0000-0002-9078-9070</orcidid><orcidid>https://orcid.org/0000-0003-3044-5388</orcidid><orcidid>https://orcid.org/0000-0001-7773-5003</orcidid><orcidid>https://orcid.org/0000-0002-9078-9070</orcidid><orcidid>https://orcid.org/0000-0001-7773-5003</orcidid><orcidid>https://orcid.org/0000-0003-2288-8277</orcidid><orcidid>https://orcid.org/0000-0003-3044-5388</orcidid><orcidid>https://orcid.org/0000-0002-6380-6349</orcidid></search><sort><creationdate>20200113</creationdate><title>Anti‐ADAMTS13 autoantibodies in immune‐mediated thrombotic thrombocytopenic purpura do not hamper ELISA‐based quantification of ADAMTS13 antigen</title><author>Dekimpe, Charlotte ; 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ADAMTS13 antigen levels are not routinely measured in iTTP patients, but studies have shown that antigen levels are a valuable prognostic factor. Objectives To (a) report the validation of our in-house developed ADAMTS13 antigen enzyme-linked immunosorbent assay (ELISA) and determine ADAMTS13 antigen in a large cohort of healthy donor and iTTP patient plasma samples; and (b) to investigate whether ADAMTS13 antigen determination is not disturbed by the presence of anti-ADAMTS13 autoantibodies.Methods Our in-house ADAMTS13 antigen ELISA was validated in terms of sensitivity, repeatability, and reproducibility. ADAMTS13 antigen levels were determined in plasma samples from 423 healthy donors and 112 acute iTTP patients. Purified IgGs from iTTP patients were added to normal human plasma to determine whether anti-ADAMTS13 autoantibodies hampered ADAMTS13 antigen determination.Results Our in-house ADAMTS13 antigen ELISA has a detection limit of 3% and low intra-assay (coefficient of variation, %CV &lt; 10%) and inter-assay (%CV &lt; 18%) variability. ADAMTS13 antigen levels were significantly reduced (P &lt; .0001) in acute iTTP patients (15 +/- 18%) compared to healthy donors (101 +/- 18%). The anti-ADAMTS13 autoantibodies in plasma of iTTP patients did not impede ADAMTS13 antigen determinations using our in-house ELISAConclusions Our in-house ADAMT13 antigen ELISA is a powerful tool to correctly determine ADAMTS13 antigen levels in iTTP patients, which supports routine ADAMTS13 antigen measurements in these patients to have better insight into disease prognosis.</abstract><pub>Wiley</pub><doi>10.1111/jth.14747</doi><orcidid>https://orcid.org/0000-0002-6380-6349</orcidid><orcidid>https://orcid.org/0000-0003-2288-8277</orcidid><orcidid>https://orcid.org/0000-0002-9078-9070</orcidid><orcidid>https://orcid.org/0000-0003-3044-5388</orcidid><orcidid>https://orcid.org/0000-0001-7773-5003</orcidid><orcidid>https://orcid.org/0000-0002-9078-9070</orcidid><orcidid>https://orcid.org/0000-0001-7773-5003</orcidid><orcidid>https://orcid.org/0000-0003-2288-8277</orcidid><orcidid>https://orcid.org/0000-0003-3044-5388</orcidid><orcidid>https://orcid.org/0000-0002-6380-6349</orcidid></addata></record>
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subjects Cardiology and cardiovascular system
Human health and pathology
Life Sciences
title Anti‐ADAMTS13 autoantibodies in immune‐mediated thrombotic thrombocytopenic purpura do not hamper ELISA‐based quantification of ADAMTS13 antigen
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