Guo Jianwei, Shen Beifen, Sun Yingxun, Yu Ming, Hu Meiru
Department of Molecular Immunology, Institute of Basic Medical Sciences, Beijing, P.R. China.
Hybridoma (Larchmt). 2006 Aug;25(4):225-9. doi: 10.1089/hyb.2006.25.225.
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.
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。
