胰蛋白酶
化学
电喷雾电离
水溶液
质谱法
肌红蛋白
色谱法
消化(炼金术)
序列(生物学)
蛋白质测序
酶
肽序列
生物化学
有机化学
基因
作者
Xiaoqin Zhong,Hao Chen,Richard N. Zare
标识
DOI:10.1038/s41467-020-14877-x
摘要
Abstract Enzymatic digestion for protein sequencing usually requires much time, and does not always result in high sequence coverage. Here we report the use of aqueous microdroplets to accelerate enzymatic reactions and, in particular, to improve protein sequencing. When a room temperature aqueous solution containing 10 µM myoglobin and 5 µg mL −1 trypsin is electrosonically sprayed (−3 kV) from a homemade setup to produce tiny (∼9 µm) microdroplets, we obtain 100% sequence coverage in less than 1 ms of digestion time, in sharp contrast to 60% coverage achieved by incubating the same solution at 37 °C for 14 h followed by analysis with a commercial electrospray ionization source that produces larger (∼60 µm) droplets. We also confirm the sequence of the therapeutic antibody trastuzumab (∼148 kDa), with a sequence coverage of 100% for light chains and 85% for heavy chains, demonstrating the practical utility of microdroplets in drug development.
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