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
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
.