{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T23:24:06Z","timestamp":1787009046774,"version":"build-2736575974"},"reference-count":16,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2025,3,26]],"date-time":"2025-03-26T00:00:00Z","timestamp":1742947200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"FWO-TBM Flanders","award":["T004821N"],"award-info":[{"award-number":["T004821N"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,7,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Anti-RuvBL1\/2 autoantibodies are found in patients with systemic sclerosis and systemic sclerosis-myositis overlap syndrome. Anti-RuvBL1\/2 antibodies recognize conformational epitopes of the RuvBL1\/2 complex, which complicates detection by solid phase assays. Here, we propose a method for detection of anti-RuvBL1\/2 autoantibodies based on liquid phase immunoprecipitation combined with quantification of the precipitated RuvBL1\/2 by LC-MS\/MS.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Serum antibodies of patients (n = 13) and controls (n = 60) were captured by and cross-linked to protein A\/G magnetic beads and incubated with HeLa nuclear cell extract. Captured proteins were eluted and digested with trypsin. The digested antigen peptides were analyzed by liquid chromatography linked to a triple quadrupole mass spectrometer operating in multiple reaction monitoring (MRM) acquisition mode. Detected signals (peak areas) were quantified. Synthetic proteotypic peptides were used for quality control and quantification purposes.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The MRM analysis was based on 6 proteotypic peptides (3 for RuvBL1, 3 for RuvBL2) that are unique to each protein. The immunoprecipitation LC-MS\/MS MRM approach allowed differentiation between anti-RuvBL1\/2 positive and anti-RuvBL1\/2 negative samples.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>We propose an immunoprecipitation-based LC-MS\/MS MRM method for anti-RuvBL1\/2 autoantibody testing. This technology is a promising detection method for autoantibodies targeting conformational epitopes.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/jalm\/jfaf072","type":"journal-article","created":{"date-parts":[[2025,5,5]],"date-time":"2025-05-05T07:42:54Z","timestamp":1746430974000},"page":"820-832","source":"Crossref","is-referenced-by-count":1,"title":["Detection of Anti-RuvBL1\/2 Autoantibodies by Targeted Liquid Chromatography\u2013Tandem Mass Spectrometry"],"prefix":"10.1093","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4572-9383","authenticated-orcid":false,"given":"Tom","family":"Dehaemers","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, University Hospitals Leuven , Leuven ,","place":["Belgium"]},{"name":"Department of Microbiology, Immunology and Transplantation, Clinical and Diagnostic Immunology, KU Leuven , Leuven ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birthe","family":"Michiels","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, University Hospitals Leuven , Leuven ,","place":["Belgium"]},{"name":"Department of Microbiology, Immunology and Transplantation, Clinical and Diagnostic Immunology, KU Leuven , Leuven ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rita","family":"Derua","sequence":"additional","affiliation":[{"name":"Laboratory for Protein Phosphorylation and Proteomics, KU Leuven , Leuven ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yves","family":"Piette","sequence":"additional","affiliation":[{"name":"Internal Medicine, Ghent University , Ghent ,","place":["Belgium"]},{"name":"Rheumatology, Ghent University Hospital , Ghent ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolien","family":"Bonroy","sequence":"additional","affiliation":[{"name":"Diagnostic Sciences, Ghent University , Ghent ,","place":["Belgium"]},{"name":"Laboratory Medicine, Ghent University Hospital , Ghent ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonas","family":"De Leeuw","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Transplantation, Clinical and Diagnostic Immunology, KU Leuven , Leuven ,","place":["Belgium"]},{"name":"Department of Rheumatology, University Hospitals Leuven , Leuven ,","place":["Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doreen","family":"Dillaerts","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Transplantation, Clinical and Diagnostic Immunology, KU Leuven , Leuven 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