Production of monoclonal antibodies against a major purgative component, sennoside B, their characterization and use in ELISA
Production of monoclonal antibodies against a major purgative component, sennoside B, their characterization and use in ELISA
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DOI：
10.1039/b103153h
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发表时间：
2001-08-01
期刊：
影响因子：
4.2
通讯作者：
Shoyama, Y
中科院分区：
文献类型：
--
作者：
Morinaga, O;Nakajima, S;Shoyama, Y
为了免疫，番泻苷B与牛血清白蛋白缀合。通过基质辅助激光解吸电离TOF质谱法测定抗原缀合物中的半抗原密度为3 mol mol（-1）蛋白质。将番泻苷B偶联物免疫小鼠，取脾细胞与小鼠骨髓瘤细胞融合，获得分泌抗番泻苷B单克隆抗体的杂交瘤细胞株。与番泻苷A（立体化学异构体）和番泻苷B的单体大黄酸发生弱交叉反应，但与其他相关蒽醌和酚类化合物未观察到交叉反应。检测范围为0.5 ng/ml ~ 15 ng/ml番泻苷B，在分析大黄粗提物时，ELISA法和HPLC法具有良好的相关性。
For immunization, sennoside B was conjugated with bovine serum albumin. The hapten density in the antigen conjugate was determined to be 3 mol mol(-1) protein by matrix-assisted laser desorption-ionization TOF mass spectrometry. A hybridoma secreting monoclonal antibody against sennoside B was produced by fusing splenocytes from mouse immunized with the sennoside B conjugate and mouse myeloma cells. Weak cross-reactivities occurred with sennoside A which is a stereochemical isomer, and a monomer of sennoside B, rhein, but no cross-reactivity was observed with other related anthraquinones and phenolics. The range of the assay extended from 0.5 ng ml(-1) to 15 ng ml(-1) of sennoside B, and good correlation between ELISA and HPLC methods was obtained when crude extracts of rhubarb were analyzed.