{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T13:57:34Z","timestamp":1791122254998,"version":"4.1.0"},"reference-count":0,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"1","license":[{"start":{"date-parts":[[2015,9,1]],"date-time":"2015-09-01T00:00:00Z","timestamp":1441065600000},"content-version":"tdm","delay-in-days":7001,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1996,7]]},"abstract":"<jats:sec>\n            <jats:title\/>\n            <jats:p>\n                                 <jats:bold>The detection of antimitochondrial autoantibodies (AMAs) is critical in the diagnosis of primary biliary cirrhosis (PBC). However, conventional laboratory assays to detect AMA are dependent on the time\u2013consuming method of immunofluorescence microscopy, a method often plagued by problems of nonspecificity. AMAs react against mitochondrial autoantigens including the E2 components of the pyruvate dehydrogenase complex (PDC\u2013E2), the branched\u2013chain 2\u2013oxo\u2013acid dehydrogenase complex (BCOADC\u2013E2), and the 2\u2013oxo\u2013glutarate dehydrogenase complex (OGDC\u2013E2). Interestingly, the immunodominant epitopes of PDC\u2013E2, BCOADC\u2013E2, and OGDC\u2013E2 are all conformational lipoate binding sites, but antibodies against them do not cross\u2013react. Although 80% to 90% of sera from patients with PBC react to PDC\u2013E2, approximately 10% patients with PBC react only to BCOADC\u2013E2 and\/or OGDC\u2013E2. We have taken advantage of our epitope\u2013mapping studies of the E2 components of PDC, BCOADC, and OGDC, and constructed a \u201cdesigner\u201d hybrid clone, designated as pML\u2013MIT3, that coexpresses the immunodominant epitopes within the three distinct lipoyl domains. We examined a total of 321 sera, including 186 sera from patients with PBC, to test the immunoreactivity of pMIT3. Of 186 sera from patients with PBC, 152 sera (81.7%) reacted with recombinant fusion protein of PDC\u2013E2, whereas 171 sera (91.9%) showed positive reactivities when probed by immunoblotting against the recombinant fusion protein expressed from the pML\u2013MIT3 clone. Of 34 PBC sera that did not react with recombinant PDC\u2013E2, 18 sera contained BCOADC\u2013E2\u2013specific AMA and 1 serum possessed only OGDC\u2013E2\u2013specific AMA. We also developed an enzyme\u2013linked immunosorbent assay (ELISA), using affinity\u2013purified recombinant fusion protein of pML\u2013MIT3 clone as the antigen source, to quantify specific AMAs in patients with PBC. None of the 135 control sera from patients with primary sclerosing cholangitis (PSC), chronic autoimmune hepatitis (CAH), systemic lupus erythematosus (SLE), or healthy volunteers showed significant reactivity against pML\u2013MIT3 recombinant fusion protein in the ELISA assay. Our results indicate that an ELISA using recombinant, cloned autoantigen of pML\u2013MIT3 is a powerful and very specific method for the detection of AMA.<\/jats:bold>\n                              <\/jats:p>\n          <\/jats:sec>","DOI":"10.1002\/hep.510240117","type":"journal-article","created":{"date-parts":[[2004,11,11]],"date-time":"2004-11-11T23:54:49Z","timestamp":1100217289000},"page":"97-103","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":121,"title":["Use of A Designer Triple Expression Hybrid Clone for Three Different Lipoyl Domain for the Detection of Antimitochondrial Autoantibodies"],"prefix":"10.1097","volume":"24","author":[{"given":"S","family":"Moteki","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P S","family":"Leung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R L","family":"Coppel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E R","family":"Dickson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M M","family":"Kaplan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S","family":"Munoz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M E","family":"Gershwin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","container-title":["Hepatology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fhep.510240117","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.lww.com\/01515467-199607000-00017","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,1]],"date-time":"2024-12-01T11:16:41Z","timestamp":1733051801000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.lww.com\/01515467-199607000-00017"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1996,7]]},"references-count":0,"journal-issue":{"issue":"1","published-print":{"date-parts":[[1996]]}},"URL":"https:\/\/doi.org\/10.1002\/hep.510240117","relation":{},"ISSN":["0270-9139"],"issn-type":[{"value":"0270-9139","type":"print"}],"subject":[],"published":{"date-parts":[[1996,7]]}}}