Foodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Services, United States Department of Agriculture, 800 Buchanan Street, Albany, CA 94710, USA.
United States Department of Homeland Security, Washington, DC 20528, USA.
Toxins (Basel). 2017 Oct 18;9(10):328. doi: 10.3390/toxins9100328.
Abrin, one of the most highly potent toxins in the world, is derived from the plant, . Because of its high toxicity, it poses potential bioterror risks. Therefore, a need exists for new reagents and technologies that would be able to rapidly detect abrin contamination as well as lead to new therapeutics. We report here a group of abrin-specific monoclonal antibodies (mAbs) that recognize abrin A-chain, intact A-B chain toxin, and agglutinin by Western blot. Additionally, these mAbs were evaluated for their ability to serve as capture antibodies for a sandwich (capture) ELISA. All possible capture-detector pairs were evaluated and the best antibody pair identified and optimized for a capture ELISA. The capture ELISA based on this capture-detector mAb pair had a limit of detection (L.O.D) of ≈1 ng/mL measured using three independent experiments. The assay did not reveal any false positives with extracts containing other potential ribosome-inactivating proteins (RIPs). Thus, this new capture ELISA uses mAbs for both capture and detection; has no cross-reactivity against other plant RIPs; and has a sensitivity comparable to other reported capture ELISAs using polyclonal antibodies as either capture or detector.
相思豆毒素,一种世界上最具毒性的毒素之一,来源于植物。由于其毒性极高,因此存在着潜在的生物恐怖风险。因此,需要新的试剂和技术来快速检测相思豆毒素的污染,并开发新的治疗方法。我们在这里报告了一组相思豆毒素特异性的单克隆抗体(mAbs),这些 mAbs 通过 Western blot 识别相思豆毒素 A 链、完整的 A-B 链毒素和凝集素。此外,还评估了这些 mAbs 作为夹心(捕获)ELISA 的捕获抗体的能力。评估了所有可能的捕获-检测对,并确定并优化了最佳的捕获 ELISA 的抗体对。基于该捕获-检测 mAb 对的捕获 ELISA 的检测限(L.O.D)约为 ≈1ng/mL,通过三个独立的实验进行测量。该检测方法在使用其他潜在核糖体失活蛋白(RIP)的提取物时没有显示任何假阳性。因此,这种新的捕获 ELISA 既使用 mAbs 进行捕获,又使用 mAbs 进行检测;对其他植物 RIPs 没有交叉反应;并且与其他使用多克隆抗体作为捕获或检测的报道的捕获 ELISA 相比,具有相当的灵敏度。