Single-day protein LC–MS bioanalysis: can next-generation trypsins cut it?: supplementary table 1

Aims: The drive toward more sensitive LC–MS assays has resulted in long, complex methods. We assessed next-generation trypsins to identify a suitable candidate to integrate into protein LC–MS method development strategies, to simplify methods and increase throughput. Materials & methods: The performance of commercially available next-generation trypsins was assessed based on the digestion of protein standards in buffer and complex matrix by LC–high-resolution MS. Results: The performance of all next-generation trypsins assessed exceeded that of an overnight tryptic digest in a fraction of the time. Performing reduction and alkylation prior to digestion with heat-stable trypsins may be beneficial and should be investigated. Conclusion: Promega Rapid-Digestion

研究目的：对灵敏度更高的液相色谱-质谱联用（LC-MS）分析方法的追求，催生了流程冗长且复杂度较高的实验方案。本研究针对新一代胰蛋白酶（trypsin）开展评估，以期筛选出可适配蛋白质LC-MS方法开发策略的理想酶种，从而简化实验流程并提升分析通量。
材料与方法：本研究通过液相色谱-高分辨质谱（LC-HRMS），基于缓冲液与复杂基质中蛋白质标准品的酶解反应，对商业化可得的新一代胰蛋白酶的性能进行评估。
结果：所有受试的新一代胰蛋白酶，均可在极短的反应时间内实现优于传统过夜胰蛋白酶酶解的实验效果。在使用热稳定型胰蛋白酶进行酶解前，先开展还原烷基化处理或可优化实验结果，值得进一步研究探索。
结论：普洛麦格（Promega）Rapid-Digestion