1. Academic Validation
  2. Ultrafast enzymatic digestion of proteins by microdroplet mass spectrometry

Ultrafast enzymatic digestion of proteins by microdroplet mass spectrometry

  • Nat Commun. 2020 Feb 26;11(1):1049. doi: 10.1038/s41467-020-14877-x.
Xiaoqin Zhong 1 Hao Chen 2 Richard N Zare 3
Affiliations

Affiliations

  • 1 Department of Chemistry, Fudan University, Shanghai, 200438, China.
  • 2 Department of Chemistry and Environmental Science, New Jersey Institute of Technology, Newark, NJ, 07102, USA.
  • 3 Department of Chemistry, Fudan University, Shanghai, 200438, China. rnz@fudan.edu.cn.
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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