Production of a Monoclonal Antibody for Sensitive Monitoring of Deoxycholic Acid Residues Anchored on Endogenous Proteins
Production of a Monoclonal Antibody for Sensitive Monitoring of Deoxycholic Acid Residues Anchored on Endogenous Proteins
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用于灵敏监测内源蛋白上脱氧胆酸残基的单克隆抗体的生产
DOI：
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
J. Goto
中科院分区：
文献类型：
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作者：
N. Kobayashi;H. Katayama;M. Nagata;J. Goto
胆汁酸羧基连接的葡萄糖醛酸酯和腺苷酸酯具有活性的酯基，因此很容易与内源性组织蛋白上的氨基发生反应，导致生理紊乱。为了在体内鉴定这种组织结合胆汁酸，我们产生了一种单克隆抗体，该抗体可以识别锚定在内源性蛋白质上的脱氧胆酸（DCA）残基。用DCA和牛血清白蛋白（BSA）偶联物免疫BALB/c小鼠脾细胞，与P3/NS1/1-Ag4-1骨髓瘤融合，生成分泌抗体的杂交瘤克隆。所得单克隆抗体（Ab#88；同型γ 1， λ）对N - ε -脱氧赖氨酸和未修饰的DCA特异，并且能够灵敏地检测由DCA腺苷酸反应引入的锚定在BSA分子上的DCA残基（270飞图）。固定Ab#88的免疫亲和柱允许选择性地提取DCA引入的物质P.该抗体将用于监测组织结合的DCA的形成和定位，这可能与DCA作为结肠癌启动子的活性有关。
Bile acid carboxy-linked glucuronides and adenylates have active ester groups, and thus may easily react with amino groups on endogenous tissue proteins resulting in physiological disorders. To identify such tissue-bound bile acids in vivo , we generated a monoclonal antibody which recognizes deoxycholic 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 antibody-secreting hybridoma clones. The resulting monoclonal antibody (Ab#88; isotype γ 1, λ ) was specific to N - ε -deoxycholyl lysine as well as the nonamidated DCA, and enabled sensitive detection of DCA residues (270 femtograms) anchored on BSA molecules, introduced by the reaction of the DCA adenylate. The immunoaffinity column immobilizing the Ab#88 allowed selective extraction of the DCA-introduced substance P. The antibody will be useful for monitoring the formation and localization of tissue-bound DCA, which may be concern with the activity of DCA as a colon cancer promoter.
石胆酸羧基连接的葡萄糖苷酸的生物合成和化学合成。
DOI：
10.1016/0005-2760(92)90294-6
发表时间：
1992
期刊：
Biochimica et biophysica acta
影响因子：
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作者：
Panfil,I;Lehman,PA;Zimniak,P;Ernst,B;Franz,T;Lester,R;Radominska,A
通讯作者：
Radominska,A
结肠癌和结肠息肉患者的组织胆汁酸。
DOI：
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发表时间：
1982
期刊：
The American journal of gastroenterology
影响因子：
--
作者：
Gelb,AM;McSherry,CK;Sadowsky,JR;Mosbach,EH
通讯作者：
Mosbach,EH
串联质谱法应用于蛋白质结构问题。
影响因子：
3.9
作者：
Biemann,K
通讯作者：
Biemann,K