Definition of antigen specificity for antimitochondrial proteins detected by Western blotting using native mitochondrial proteins in primary biliary cirrhosis

Citation
H. Miyakawa et al., Definition of antigen specificity for antimitochondrial proteins detected by Western blotting using native mitochondrial proteins in primary biliary cirrhosis, HEPATOL RES, 21(2), 2001, pp. 101-107
Citations number
32
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Gastroenerology and Hepatology
Journal title
HEPATOLOGY RESEARCH
ISSN journal
1386-6346 → ACNP
Volume
21
Issue
2
Year of publication
2001
Pages
101 - 107
Database
ISI
SICI code
1386-6346(200110)21:2<101:DOASFA>2.0.ZU;2-8
Abstract
The major autoantigens to anti-mitochondrial antibody (AMA) in primary bili ary cirrhosis (PBC) have previously been identified to be PDC-E2, BCOADC-E2 , and OGDC-E2. However, analysis of these autoantigens to AMA cannot be exa mined using the two routine assays; immmunofluorescence and ELISA. Moreover , there are some problems in specificity and sensitivity in these routine a ssays. So, analysis with Western blotting using native mitochondrial protei n as the antigen is required; it allows the identification of the molecular weights for the proteins which react with AMA in patients' sera. However, since the antigen-proteins used are not unified, molecular weights of AMA c orresponding proteins vary among laboratories. In the present study, as the first step to help address this issue, we investigated the antigen specifi city of protein bands detected by Western blotting using our in-house bovin e and porcine heart mitochondrial proteins, Three major recombinant mitocho ndrial proteins were prepared. The antigen specificity was examined by the absorption tests preincubated with the three recombinant mitochondrial prot eins. The molecular weights of developing our bovine and porcine heart mito chondrial proteins using SDS-PAGE were multiple protein bands including 74, 52, 50, and 43 kDa. protein bands. Of them, the 74, 50, and 43 kDa protein bands were absorbed with preincubations of recombinant PDC-E2, BCOADC-E2, and OGDC-E2 protein, respectively. AMA specificity of these three major pro teins with our Western blotting was confirmed. (C) 2001 Elsevier Science B. V. All rights reserved.