MRM quantification of circulating CFP-10 in TB patients

Liquid chromatography coupled mass spectrometry (LC-MS) operating in targeted mass spectrometry (MS) including multiple reaction monitoring (MRM) mode is an accurate analytical method. Integrating with nanoelectrospray-tandem MS (NanoES-MS/MS) and immunoprecipitation (IP), it becomes a promising method in detecting pathogen derived antigens which are at extremely low concentrations, such as mycobacterium tuberculosis (Mtb) secreted 10 kDa culture filtrate antigen (CFP-10). We employed an optimum denaturing condition for trypsin digestion and imunoprecipitation of targeted peptides and applied this method in clinical samples.液相色谱-质谱联用（Liquid chromatography coupled mass spectrometry, LC-MS）在靶向质谱（targeted mass spectrometry, MS）模式下运行，涵盖多反应监测（multiple reaction monitoring, MRM）模式，是一种精准可靠的分析方法。将其与纳米电喷雾串联质谱（nanoelectrospray-tandem MS, NanoES-MS/MS）及免疫沉淀（immunoprecipitation, IP）技术联用后，该方法可用于检测浓度极低的病原体源性抗原，例如结核分枝杆菌（mycobacterium tuberculosis, Mtb）分泌的10 kDa培养滤液抗原（CFP-10），成为极具应用潜力的检测手段。本研究优化了目标肽段胰蛋白酶消化与免疫沉淀的最佳变性条件，并将该方法应用于临床样本检测。