毒性机制导向式SPR-HRMS：活性生物毒素的一体化高灵敏定量及结构区分鉴定研究

针对防化医学、军事分析化学方面的迫切需求，本项目拟结合生物质谱和亲和技术，创新性提出“以毒性机制为导向，结合多种纳米功能化亲和元件，于SPR-HRMS中，同时实现活性毒素蛋白的一体化识别、定量检测及准确结构鉴定”的科学思路，以SPR提供高灵敏、实时生物分子互作及定量信息，以纳米功能化亲和元件提供特异性识别、双重信号放大、高效发挥毒素分析物的糖苷酶活性，以及以HRMS(MS)提供精准的结构鉴定功能，从而重点解决活性蓖麻毒素与灭活毒素蛋白区分及高灵敏准确定量、活性测定、以及结构相似毒素蛋白的精确结构区分鉴定的关键难点问题。并将该方案用于实际复杂基质中活性毒素的成功检测，及探讨在新改构毒素、其它酶催化类型毒素中的拓展应用，为防化分析、中毒诊断、防护救治等领域提供重要的科学技术支撑。

In response to the urgent needs of prevention medicine on chemical warfare agents and military analytical chemistry, this project intends to combine biological mass spectrometry and affinity techniques, and innovatively proposes the concept of “a toxicity mechanism-oriented integrated realization of identification, quantitative detection and accurate structural identification towards active toxin proteins, in combination with multiple functionalized nano-affinity material by SPR-HPMS techniques”. Here we provide highly sensitive, real-time biomolecular interactions and quantitative information with SPR, offer specific recognition, dual signal amplification, and efficient fulfillment of the N-glycosidase activity of toxin analytes with functionalized nano-affinity material, and provide accurate structural identification with high resolution tandem mass spectrometry, HRMS(MS). Therefore, it can address the key problem of distinguishing active ricin toxin from inactivated toxin protein, with highly sensitive, accurate quantification and activity determination, as well as precisely structural identification of structurally similar toxin protein. The scheme can then be applied to the successful detection of toxin molecules in the real complicated matrix, and explores the expansion and application of newly modified toxins and other types of toxins with enzymatic characteristics. It will provide important scientific and technical basis for the fields of analysis, diagnosis, protection and treatment of chemical warfare agents.