{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T12:14:20Z","timestamp":1768306460299,"version":"3.49.0"},"reference-count":21,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T00:00:00Z","timestamp":1768262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Aim<\/jats:title>\n                    <jats:p>Neutralizing anti-drug antibodies (ADA) limit therapy efficacies significantly. Pegunigalsidase alfa is a newly approved drug for the treatment of Fabry disease. The increased plasma half-life due to PEGylation interferes with ADA measurements including ELISAs and serum-mediated inhibition assays. We developed a rapid protocol eliminating pegunigalsidase alfa from blood samples, without interfering downstream applications.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A rapid protocol based on alkaline-pretreatment followed by acid-based neutralization of agalsidase-spiked control sera was established. Results were confirmed using serum samples from patients with and without neutralizing ADAs drawn during infusions. Repeated ADA measurements including serum-mediated inhibition assays and ELISA-based immunoglobulin isotyping (IgG, IgA, IgM) were performed with sera from 17 patients receiving pegunigalsidase alfa.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Alkaline pretreatment with NaOH was sufficient to eliminate up to 1 \u00b5g\/ml agalsidase alfa or pegunigalsidase alfa in control sera. AGAL activities in sera drawn during infusions were completely suppressed without interfering subsequent serum-mediated inhibition assays. Based on this method, in one patient a\n                      <jats:italic>de novo<\/jats:italic>\n                      formation of ADAs against pegunigalsidase alfa was identified. Immunoglobulin isotyping showed mainly IgM antibodies towards pegunigalsidase alfa, recognizing PEG moieties and amino acids in this patient. Although his ADAs had a low inhibitory capacity, Western blot analyses demonstrated that the reduced pharmacokinetics might be linked to leucocyte-mediated enzyme elimination. A second patient with pre-existing ADAs before pegunigalsidase alfa-initiation showed a massive induction of anti-PEG antibodies with inhibitory function.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>We present a rapid alkaline-treatment based method to overcome drug interferences to measure at least free antibodies in patients treated with pegunigalsidase alfa.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2025.1724835","type":"journal-article","created":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T06:35:16Z","timestamp":1768286116000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A rapid method to reduce drug interferences for antibody measurements in pegunigalsidase alfa-treated patients with Fabry disease"],"prefix":"10.3389","volume":"16","author":[{"given":"Malte","family":"Lenders","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Rudolph","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Rudnicki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Markus","family":"Cybulla","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Brand","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(08)61589-5","article-title":"Fabry\u2019s 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neutralizing anti-drug antibody formation and clinical relevance on therapeutic efficacy of enzyme replacement therapies in Fabry disease","volume":"81","author":"Lenders","year":"2021","journal-title":"Drugs"},{"key":"B12","doi-asserted-by":"publisher","first-page":"10661","DOI":"10.1021\/acs.analchem.6b03109","article-title":"Measurement of pre-existing IgG and IgM antibodies against polyethylene glycol in healthy individuals","volume":"88","author":"Chen","year":"2016","journal-title":"Anal Chem"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1266082","article-title":"Characterization of pre-existing anti-PEG and anti-AGAL antibodies towards PRX-102 in patients with Fabry disease","volume":"14","author":"Lenders","year":"2023","journal-title":"Front Immunol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-024-00690-1","article-title":"Relevance of neutralizing antibodies for the pharmacokinetics of pegunigalsidase alfa in patients with 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