Pauly J U, Bitter-Suermann D, Dose K
Institut für Medizinische Mikrobiologie, Universität Mainz.
Biol Chem Hoppe Seyler. 1988 Jun;369(6):487-92. doi: 10.1515/bchm3.1988.369.1.487.
A monoclonal antibody was obtained by the fusion of mouse myeloma cells with splenocytes isolated from Balb/c mice, which had been immunized with diacetoxyscirpenol-hemiglutarate (DAS-hemiglutarate) and verrucarol-hemiglutarates covalently bound to ethylenediamine-modified bovine serum albumin. The anti-DAS-antibody that could be induced was of the IgM type with kappa-chains. The titer of the monoclonal anti-DAS-antibody in ascites fluid obtained from mice injected the selected cell line was much higher than those of conventional antisera. An enzyme-linked immunosorbent assay based on the competitive binding principle in which the antibody was applied had a sensitivity of 1 ng DAS per assay. The relative cross-reactivity of the monoclonal antibody in the CI-ELISA with the related trichothecenes such as triacetoxyscirpenol, 15-monoacetoxyscirpenol, diacetylverrucarol, 4-monoacetoxyscirpenol and scirpentriol were found to be 1.8, 0.8, 0.15, 0.02 and less than 0.001, respectively. The trichothecenes verrucarol, T-2 toxin, T-2 tetraol, deoxynivalenol, 3-acetyldeoxynivalenol and trichothecin showed no cross-reactivity.
通过将小鼠骨髓瘤细胞与从经二乙酰氧基雪腐镰刀菌烯醇-半谷氨酸酯(DAS-半谷氨酸酯)和疣孢菌素-半谷氨酸酯共价结合到乙二胺修饰的牛血清白蛋白免疫的Balb/c小鼠中分离的脾细胞融合,获得了一种单克隆抗体。可诱导产生的抗DAS抗体为κ链IgM型。从注射了所选细胞系的小鼠获得的腹水中单克隆抗DAS抗体的效价比传统抗血清高得多。基于竞争结合原理应用该抗体的酶联免疫吸附测定法的灵敏度为每次测定1 ng DAS。发现该单克隆抗体在CI-ELISA中与相关单端孢霉烯族毒素如三乙酰氧基雪腐镰刀菌烯醇、15-单乙酰氧基雪腐镰刀菌烯醇、二乙酰疣孢菌素、4-单乙酰氧基雪腐镰刀菌烯醇和蛇形菌素的相对交叉反应率分别为1.8、0.8、0.15、0.02和小于0.001。单端孢霉烯族毒素疣孢菌素、T-2毒素、T-2四醇、脱氧雪腐镰刀菌烯醇、3-乙酰脱氧雪腐镰刀菌烯醇和西瓜镰孢菌素未显示交叉反应。