PDF(Pubmed)
Abstract：
Ricin and abrin are highly potent plant-derived toxins, categorized as type II ribosome-inactivating proteins. High toxicity, accessibility, and the lack of effective countermeasures make them potential agents in bioterrorism and biowarfare, posing significant threats to public safety. Despite the existence of many effective analytical strategies for detecting these two lethal toxins, current methods are often hindered by limitations such as insufficient sensitivity, complex sample preparation, and most importantly, the inability to distinguish between biologically active and inactive toxin. In this study, a cytotoxicity assay was developed to detect active ricin and abrin based on their potent cell-killing capability. Among nine human cell lines derived from various organs, HeLa cells exhibited exceptional sensitivity, with limits of detection reaching 0.3 ng/mL and 0.03 ng/mL for ricin and abrin, respectively. Subsequently, toxin-specific neutralizing monoclonal antibodies MIL50 and 10D8 were used to facilitate the precise identification and differentiation of ricin and abrin. The method provides straightforward and sensitive detection in complex matrices including milk, plasma, coffee, orange juice, and tea via a simple serial-dilution procedure without any complex purification and enrichment steps. Furthermore, this assay was successfully applied in the unambiguous identification of active ricin and abrin in samples from OPCW biotoxin exercises.
摘要：
蓖麻毒素和abrin是非常有效的植物衍生毒素，分类为II型核糖体失活蛋白。高毒性,可访问性,缺乏有效的对策使其成为生物恐怖主义和生物战的潜在代理人，对公共安全构成重大威胁。尽管存在许多检测这两种致命毒素的有效分析策略，目前的方法往往受到灵敏度不足等限制的阻碍，复杂的样品制备，最重要的是,无法区分生物活性和非活性毒素。在这项研究中,开发了一种细胞毒性测定法，以检测活性蓖麻毒素和相思毒素，基于其强大的细胞杀伤能力。在九种来自不同器官的人类细胞系中，HeLa细胞表现出异常的敏感性，蓖麻毒素和abrin的检测限达到0.3ng/mL和0.03ng/mL，分别。随后，毒素特异性中和单克隆抗体MIL50和10D8被用来促进蓖麻毒素和相思子的精确鉴定和区分。该方法在包括牛奶在内的复杂基质中提供了简单而灵敏的检测，等离子体,咖啡,橙汁,和茶通过一个简单的连续稀释程序没有任何复杂的纯化和富集步骤。此外，该方法已成功应用于对禁化武组织生物毒素实验样品中的活性蓖麻毒素和相思子蛋白的明确鉴定。
