{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T22:53:00Z","timestamp":1780527180323,"version":"3.54.1"},"reference-count":27,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,10,3]],"date-time":"2022-10-03T00:00:00Z","timestamp":1664755200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2023,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>The development of neutralising (inhibitors) and non\u2010neutralising antibodies (NNAs) is a complication to factor replacement therapy in haemophilia. The diagnostic methods available lack standardisation, have high inter\u2010laboratory variation, and false\u2010negative as well as false\u2010positive results may affect treatment. Both functional inhibitors and NNAs may be detected with higher reproducibility, sensitivity and specificity using the immunological Luminex xMAP\u2010based fluorescence\u2010immunoassay (xFLI).<\/jats:p><\/jats:sec><jats:sec><jats:title>Aim<\/jats:title><jats:p>Validation of our xFLI and comparability with enzyme\u2010linked immunosorbent assay (ELISA) and chromogenic Nijmegen\u2010Bethesda assay (CBA) for anti\u2010FVIII antibodies in haemophilia A (HA) patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The xFLI method was developed with full\u2010length and B\u2010domain deleted factor coupled to magnetic beads, optimised and validated for performance characteristics. Comparability with ELISA and CBA was evaluated in HA patient samples (<jats:italic>n<\/jats:italic>\u00a0=\u00a0112), serial samples in six inhibitor patients and reference interval and decision\u2010limits in healthy donors (<jats:italic>n<\/jats:italic>\u00a0=\u00a044).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The intra\u2010 and inter\u2010assay precision (CV%) for the xFLI method was below 6% and detection limit (LLOQ) .084\u00a0ng\/mL (NovoEight). All ELISA\u2010positive samples were positive with either Advate or NovoEight. Additionally, 10.7%\u201314.3% were xFLI\u2010positive and ELISA\u2010negative. All but one CBA\u2010positive sample was above 3SD with xFLI; one was between 2 and 3SD. 29.1% were xFLI\u2010positive and CBA negative. The overall concordance between xFLI and ELISA was 82.1% and xFLI and CBA 77.9%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The anti\u2010FVIII antibody xFLI method is adaptable to clinical practice and more sensitive and reproducible than ELISA and CBA. Actual NNA titers are determined to both full\u2010length and B\u2010domain deleted FVIII. The xFLI is thus valuable for confirmation of all anti\u2010FVIII antibodies.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hae.14669","type":"journal-article","created":{"date-parts":[[2022,10,4]],"date-time":"2022-10-04T04:59:41Z","timestamp":1664859581000},"page":"336-347","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Methods for anti\u2010factor VIII antibody levels in haemophilia A patients \u2013 validation of a multiplex immunoassay and comparability with assays measuring non\u2010neutralising and neutralising antibodies (inhibitors)"],"prefix":"10.1111","volume":"29","author":[{"given":"Myriam","family":"Martin","sequence":"first","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional Laboratories Region Sk\u00e5ne  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5587-3712","authenticated-orcid":false,"given":"Cecilia","family":"Augustsson","sequence":"additional","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional Laboratories Region Sk\u00e5ne  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivian","family":"Lind","sequence":"additional","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional Laboratories Region Sk\u00e5ne  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riam","family":"Al\u2010Sabti","sequence":"additional","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional Laboratories Region Sk\u00e5ne  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"My Chi","family":"Lam","sequence":"additional","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional Laboratories Region Sk\u00e5ne  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadine G.","family":"Andersson","sequence":"additional","affiliation":[{"name":"Department of Thrombosis and Haemostasis Hematology Oncology and Radiation Physics Sk\u00e5ne University Hospital  Malm\u00f6 Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5554-6631","authenticated-orcid":false,"given":"Karin","family":"Strandberg","sequence":"additional","affiliation":[{"name":"Department of Clinical Chemistry and Pharmacology Division of Laboratory Medicine Coagulation Laboratory University and Regional 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