Production of a monoclonal antibody for sensitive monitoring of deoxycholic acid residues anchored on endogenous proteins

Citation
N. Kobayashi et al., Production of a monoclonal antibody for sensitive monitoring of deoxycholic acid residues anchored on endogenous proteins, ANAL SCI, 16(11), 2000, pp. 1133-1138
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Spectroscopy /Instrumentation/Analytical Sciences
Journal title
ANALYTICAL SCIENCES
ISSN journal
0910-6340 → ACNP
Volume
16
Issue
11
Year of publication
2000
Pages
1133 - 1138
Database
ISI
SICI code
0910-6340(200011)16:11<1133:POAMAF>2.0.ZU;2-Z
Abstract
Bile acid carboxy-linked glucuronides and adenylates have active ester grou ps, and thus may easily react with amino groups on endogenous tissue protei ns resulting in physiological disorders. To identify such tissue-bound bile acids in vivo, we generated a monoclonal antibody which recognizes deoxych olic acid (DCA) residues anchored on endogenous proteins. The spleen cells from a BALB/c mouse, which was immunized with a conjugate of DCA and bovine serum albumin (BSA), were fused with P3/NS1/1-Ag4-1 myeloma to generate an tibody-secreting hybridoma clones. The resulting monoclonal antibody (Ab#88 ; isotype gamma1, lambda) was specific to N-epsilon -deoxycholyl lysine as well as the nonamidated DCA, and enabled sensitive detection of DCA residue s (270 femtograms) anchored on BSA molecules, introduced by the reaction of the DCA adenylate. The immunoaffinity column immobilizing the Ab#88 allowe d selective extraction of the DCA-introduced substance P. The antibody will be useful for monitoring the formation and localization of tissue-bound DC A, which may be concern with the activity of DCA as a colon cancer promoter .