Faculty of Military Health Sciences, University of Defense in Brno, Třebešská 1575, CZ-500 01 Hradec Králové, Czech Republic.
Department of Biology, Faculty of Science, University of Hradec Kralove, Hradecká 1285, CZ-500 03 Hradec Kralove, Czech Republic.
Toxins (Basel). 2019 Mar 23;11(3):177. doi: 10.3390/toxins11030177.
Targeted proteomics recently proved to be a technique for the detection and absolute quantification of proteins not easily accessible to classical bottom-up approaches. Due to this, it has been considered as a high fidelity tool to detect potential warfare agents in wide spread kinds of biological and environmental matrices. toxins are considered to be potential biological weapons, especially the epsilon toxin which belongs to a group of the most powerful bacterial toxins. Here, the development of a target mass spectrometry method for the detection of protein toxins (alpha, beta, beta2, epsilon, iota) is described. A high-resolution mass spectrometer with a quadrupole-Orbitrap system operating in target acquisition mode (parallel reaction monitoring) was utilized. Because of the lack of commercial protein toxin standards recombinant toxins were prepared within . The analysis was performed using proteotypic peptides as the target compounds together with their isotopically labeled synthetic analogues as internal standards. Calibration curves were calculated for each peptide in concentrations ranging from 0.635 to 1101 fmol/μL. Limits of detection and quantification were determined for each peptide in blank matrices.
靶向蛋白质组学最近被证明是一种检测和绝对定量经典自上而下方法不易获得的蛋白质的技术。因此，它被认为是一种高保真工具，可用于检测广泛存在于生物和环境基质中的潜在战争制剂。毒素被认为是潜在的生物武器，特别是属于最强细菌毒素之一组的ε毒素。在这里，描述了一种用于检测蛋白质毒素（α、β、β2、ε、iota）的靶向质谱方法。使用具有四极杆-Orbitrap 系统的高分辨率质谱仪以目标采集模式（平行反应监测）运行。由于缺乏商业蛋白质毒素标准品，因此在 内制备了重组毒素。分析使用作为目标化合物的蛋白肽以及它们的同位素标记的合成类似物作为内标进行。针对每个肽在 0.635 至 1101 fmol/μL 的浓度范围内计算校准曲线。在空白基质中确定每个肽的检测限和定量限。
