{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T20:49:58Z","timestamp":1791838198247,"version":"4.3.4"},"reference-count":29,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2023,10,12]],"date-time":"2023-10-12T00:00:00Z","timestamp":1697068800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2024,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Patients with hemophilia A treated with coagulation Factor VIII (FVIII) products are at risk for developing anti\u2010FVIII antibodies. The ABIRISK Consortium aimed to provide knowledge on the formation and detection of anti\u2010drug antibodies against biopharmaceutical products, including FVIII. Accordingly, standardized and validated assays for the detection of binding (total) and neutralizing antibodies are needed.<\/jats:p><\/jats:sec><jats:sec><jats:title>Aim<\/jats:title><jats:p>Two\u2010center validation of an ELISA for the detection of total FVIII\u2010binding IgG\u2010antibodies and Nijmegen\u2010Bethesda assays for the quantification of FVIII\u2010neutralizing antibodies according to consensus validation guidelines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Validation of assays at both sites was done according to published recommendations and included preanalytics, the determination of key assay parameters, including cut\u2010points, assay sensitivity, precision, and FVIII interference.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The validated assays reproducibly detected FVIII\u2010binding and \u2010neutralizing antibodies with comparable performance in both laboratories. Floating screening cut\u2010points were established for both assays. Determined mass\u2010based sensitivity of both assays (all values \u226466\u00a0ng\/mL) complied with the minimum sensitivity for the detection of anti\u2010drug antibodies as recommended by the FDA (&lt;100\u00a0ng\/mL). Intra\u2010 and inter\u2010assay coefficients of variation did not exceed 25%. Assay validation further revealed that pre\u2010analytical heat treatment led to potentially false\u2010positive ELISA results, while up to 0.15 IU\/mL, residual FVIII showed no significant impact. Overall, good agreement of results was found for patient samples analyzed at both study sites.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Comprehensive validation of different anti\u2010FVIII\u2010antibody assays in two laboratories gave novel insights into the impact of pre\u2010analytical sample treatment as well as the comparability of test results generated by the use of methodically different assays.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hae.14885","type":"journal-article","created":{"date-parts":[[2023,10,12]],"date-time":"2023-10-12T17:38:16Z","timestamp":1697132296000},"page":"224-231","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Two\u2010center validation of assays for the detection of binding and neutralizing anti\u2010factor VIII antibodies"],"prefix":"10.1111","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7713-9631","authenticated-orcid":false,"given":"Jens","family":"M\u00fcller","sequence":"first","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine University Hospital Bonn  Bonn Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonja","family":"Neimanis","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine Clinical and Molecular Hemostasis Goethe University Frankfurt University Hospital  Frankfurt am Main Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f6rg","family":"Kahle","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine Clinical and Molecular Hemostasis Goethe University Frankfurt University Hospital  Frankfurt am Main Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thilo","family":"Albert","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine University Hospital Bonn  Bonn Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephan","family":"Schultze Strasser","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine Clinical and Molecular Hemostasis Goethe University Frankfurt University Hospital  Frankfurt am Main Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bonita","family":"Rup","sequence":"additional","affiliation":[{"name":"Pfizer Immunogenicity Sciences Disciple Pharmacokinetics Dynamics and Metabolism  New York NY USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernd","family":"P\u00f6tzsch","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine University Hospital Bonn  Bonn Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph","family":"K\u00f6nigs","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine Clinical and Molecular Hemostasis Goethe University Frankfurt University Hospital  Frankfurt am Main Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1585-4100","authenticated-orcid":false,"given":"Johannes","family":"Oldenburg","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine University Hospital Bonn  Bonn Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"name":"the ABIRISK Consortium","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,10,12]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12659"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1703068"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.blre.2018.02.003"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1055\/a-1665-6232"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-0038-1653759"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.13424"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.12956"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.13565"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/jth.12672"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.12666"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.14669"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtha.2023.03.011"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2015-10-675512"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-06-720086"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.14109"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12652"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2008.09.020"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V92.2.496"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/bjh.12923"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/hae.13435"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2021005745"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(84)90445-9"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(84)90446-0"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(88)90311-0"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-08-174987"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1208\/s12248-011-9263-z"},{"key":"e_1_2_10_28_1","unstructured":"Immunogenicity Testing of Therapeutic Protein Products \u2014 Developing and Validating Assays for Anti\u2010Drug Antibody Detection. US Food and Drug Administration (FDA).2019. Docket Number: FDA\u20102009\u2010D\u20100539."},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(96)10067-6"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1111\/jth.12259"}],"container-title":["Haemophilia"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/hae.14885","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,5]],"date-time":"2024-04-05T13:08:51Z","timestamp":1712322531000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/hae.14885"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,12]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,1]]}},"alternative-id":["10.1111\/hae.14885"],"URL":"https:\/\/doi.org\/10.1111\/hae.14885","archive":["Portico"],"relation":{},"ISSN":["1351-8216","1365-2516"],"issn-type":[{"value":"1351-8216","type":"print"},{"value":"1365-2516","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,12]]},"assertion":[{"value":"2023-08-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-09-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-10-12","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}