Epsilon toxin (ETX), produced by Clostridium perfringens Type B or type D strains, is a potential biological and toxin warfare (BTW) agent, largely for its very high toxicity. The toxin is implicated in several animal diseases. Using LC-MS/MS analysis, we report here elucidation of putative serum maker proteins for ETX exposure with an objective of the early diagnosis of intoxication. Of 166 consensus proteins (488 peptides), showing ETX-induced alterations, 119 proteins exhibited increase and 47 proteins showed decreased abundance in serum, as revealed by SWATH (DIA) acquisition on LC-MS/MS and label free quantitative analysis of control and test samples. Complement and coagulation cascade, nitrogen metabolism, negative regulation of peptidase activity, and response to ROS were among the biological processes and pathways perturbed by the ETX exposure. Interaction network indicated enzyme inhibitor activity, detoxification of ROS, and steroid binding functions were the major interaction networks for the proteins with increased abundance, while, hemostasis and structural molecule activity were the prominent networks for the down-regulated proteins. Validation studies were carried out by immunoprecipitation, ELISA, and Western blot analysis of selected proteins to demonstrate diagnostic potential of the putative marker proteins of ETX exposure.

产气荚膜梭菌产生的Epsilon毒素（ETX）B型或D型菌株是一种潜在的生物和毒素战（BTW）药剂，主要是因为它的毒性很高。该毒素与几种动物疾病有关。使用LC-MS / MS分析，我们在这里报告了ETX暴露的假定血清制造者蛋白的阐明，目的是早期诊断中毒。在LC-MS / MS上通过SWATH（DIA）采集以及对照和对照的无标记定量分析表明，在166个共有蛋白（488个肽）中，它们显示出ETX诱导的改变，其中119个蛋白表现出增加，47个蛋白显示出降低的血清丰度。测试样品。补体和凝血级联反应，氮代谢，肽酶活性的负调控以及对ROS的反应都是受到ETX暴露干扰的生物学过程和途径。相互作用网络表明酶抑制剂的活性，ROS的解毒作用和类固醇结合功能是丰度增加的蛋白质的主要相互作用网络，而止血和结构分子活性是下调蛋白质的主要网络。通过免疫沉淀，ELISA和选定蛋白的蛋白质印迹分析进行了验证研究，以证明ETX暴露的假定标记蛋白的诊断潜力。