{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T12:53:55Z","timestamp":1791204835353,"version":"4.1.0"},"reference-count":40,"publisher":"Wiley","issue":"S1","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100018748","name":"Argenx","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100018748","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Muscle and Nerve"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Introduction\/Aims<\/jats:title>\n                    <jats:p>Degradation of cell\u2010surface acetylcholine receptors (AChRs) by antigenic modulation is a key pathogenic mechanism of myasthenia gravis (MG) autoantibodies, yet standard assays primarily detect antibody binding rather than functional effects. We therefore aimed to establish a standardized assay to evaluate the functional effects of MG autoantibodies on AChR modulation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      CN21 cells expressing adult and fetal\u2010type AChR were used to measure the modulation capacity of AChR antibodies in\u00a0vitro. In this cell\u2010based assay, the remaining surface AChR levels were determined using iodine\u2010125\u2013conjugated\n                      <jats:italic>\u03b1<\/jats:italic>\n                      \u2010bungarotoxin. Experimental variables including plate surface and coating, cell density, dexamethasone treatment, incubation time, and complement effect were systematically evaluated and optimized. The optimized assay was later tested and qualified using sera from AChR\u2010MG patients and healthy individuals.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>CellBIND coated 48\u2010well plates were selected because of robust cell adhesion. An increased cell density resulted in increased baseline AChR expression levels without affecting the modulation capacity. Prolonged incubation of cells with AChR antibodies for 3\u2009h resulted in higher sensitivity compared with 1\u2009h. Heat inactivation of complement in serum samples did not affect antigenic modulation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>The optimized modulation assay showed strong sensitivity and specificity, and is a robust and reproducible method to quantify modulation capacity in the sera of individual AChR\u2010MG patients. We encourage the use of this assay to characterize the relevance of modulating antibodies in clinical severity and treatment response in individual patients.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/mus.70319","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T04:58:59Z","timestamp":1782968339000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimization of the Cell\u2010Based Assay Methodology to Study Degradation of Acetylcholine Receptors (Antigenic Modulation) by Antibodies From Myasthenia Gravis Patients"],"prefix":"10.1002","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-0298-1493","authenticated-orcid":false,"given":"Peng","family":"Du","sequence":"first","affiliation":[{"name":"Department of Psychiatry and Neuropsychology Mental Health and Neuroscience Research Institute, Faculty of Health, Medicine and Life Sciences, Maastricht University  Maastricht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Man\u00e9\u2010Damas","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology Mental Health and Neuroscience Research Institute, Faculty of Health, Medicine and Life Sciences, Maastricht University  Maastricht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fien M.","family":"Verhamme","sequence":"additional","affiliation":[{"name":"argenx  Ghent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahan","family":"Moshir","sequence":"additional","affiliation":[{"name":"argenx  Ghent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Ulrichts","sequence":"additional","affiliation":[{"name":"argenx  Ghent Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbie","family":"Machiels","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology Mental Health and Neuroscience Research Institute, Faculty of Health, Medicine and Life Sciences, Maastricht University  Maastricht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9761-0002","authenticated-orcid":false,"given":"Mario","family":"Losen","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology Mental Health and Neuroscience Research Institute, Faculty of Health, Medicine and Life Sciences, Maastricht University  Maastricht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5990-3373","authenticated-orcid":false,"given":"Pilar","family":"Martinez\u2010Martinez","sequence":"additional","affiliation":[{"name":"Department of Psychiatry and Neuropsychology Mental Health and Neuroscience Research Institute, Faculty of Health, Medicine and Life Sciences, Maastricht University  Maastricht the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-019-0079-y"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.autrev.2025.103875"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrneurol.2016.44"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0025-6196(25)09826-X"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.29.2.179"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/mus.880020304"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.28.2.130"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(17)30369-1"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2023.1284444"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.144.3.739"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)50366-5"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM197805182982004"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.132.2.711"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/267263a0"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1097\/00005072-197609000-00008"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.74.5.2130"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.850793"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.74.7.3090"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/274065a0"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/274068a0"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.663610"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2825(1998)12:5<315::AID-JCLA12>3.0.CO;2-Y"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/0896-8411(89)90004-8"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/0304-3940(96)12488-5"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/eji.200425671"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.136.9.3231"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-01019-5"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.158.4.1919"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.4c03213"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1309\/AJCPYEOR6SGE8ZLU"},{"key":"e_1_2_12_32_1","doi-asserted-by":"publisher","DOI":"10.1212\/NXI.0000000000200436"},{"key":"e_1_2_12_33_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00401-023-02603-y"},{"key":"e_1_2_12_34_1","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.0000000000010910"},{"key":"e_1_2_12_35_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.20.8100"},{"key":"e_1_2_12_36_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.1993.tb13422.x"},{"key":"e_1_2_12_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/jnr.490310209"},{"key":"e_1_2_12_38_1","doi-asserted-by":"publisher","DOI":"10.1196\/annals.1405.034"},{"key":"e_1_2_12_39_1","doi-asserted-by":"publisher","DOI":"10.1002\/1531-8249(200011)48:5<696::AID-ANA2>3.0.CO;2-S"},{"key":"e_1_2_12_40_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.00097"},{"key":"e_1_2_12_41_1","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12217"}],"container-title":["Muscle 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