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一种针对蓖麻毒素A和蓖麻毒素B的新型中和单克隆抗体及其快速夹心酶联免疫吸附测定法的应用。

A novel neutralizing monoclonal antibody against both ricin toxin A and ricin toxin B, and application of a rapid sandwich enzyme-linked immunosorbent assay.

作者信息

Guo Jianwei, Shen Beifen, Sun Yingxun, Yu Ming, Hu Meiru

机构信息

Department of Molecular Immunology, Institute of Basic Medical Sciences, Beijing, P.R. China.

出版信息

Hybridoma (Larchmt). 2006 Aug;25(4):225-9. doi: 10.1089/hyb.2006.25.225.

Abstract

Two strains of neutralizing monoclonal antibodies (MAbs) anti-ricin, both B chain and A chain, named 3D74 and 4C13, were generated efficiently. The two antibodies recognized different epitopes located in a separated toxin structure domain characterized by enzyme-linked immunosorbent assay (ELISA) and Western blotting. 3D74 recognized space conformation epitope, whereas 4C13 recognized linearity epitope of ricin. 4C13 possessed a novel neutralizing ability; the safe period for intraperitoneal injection of 100 microg of antibody was 30 min after intraperitoneal injection of 2 microg of ricin (10 times LD50). Using 3D74 and 4C13 as pair-matching antibodies, we established sandwich ELISA and rapid ELISA for detection of ricin. The detection sensitivities were 31.3 and 156 pg/ml, respectively. Rapid sandwich ELISA was sufficiently sensitive and was performed easily for ricin detection without any special instruments.

摘要

高效制备了两株抗蓖麻毒素的中和单克隆抗体(MAb),分别针对B链和A链,命名为3D74和4C13。通过酶联免疫吸附测定(ELISA)和蛋白质印迹法鉴定,这两种抗体识别位于分离毒素结构域中的不同表位。3D74识别空间构象表位,而4C13识别蓖麻毒素的线性表位。4C13具有一种新的中和能力;腹腔注射2μg蓖麻毒素(10倍半数致死量)后,腹腔注射100μg抗体的安全期为30分钟。以3D74和4C13作为配对抗体,建立了检测蓖麻毒素的夹心ELISA和快速ELISA方法。检测灵敏度分别为31.3和156 pg/ml。快速夹心ELISA灵敏度足够高,且无需任何特殊仪器即可轻松用于蓖麻毒素检测。

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