Chymotryptic proteolysis accelerated by alternating current for MALDI-TOF-MS peptide mapping
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Updated: Jun 26, 2026

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor
Published on: April 6, 2016
Sheng Wang1, Ting Liu, Luyan Zhang
1Department of Chemistry, School of Pharmacy, Fudan University, Shanghai, China.
Abstract:
Alternating current (AC) has been employed to enhance the efficiency of chymotryptic proteolysis for peptide mapping. It was allowed to flow through the mixture solution of proteins and chymotrypsin via a pair of platinum wire electrodes. Bovine serum albumin (BSA) and cytochrome c (Cyt-c) were digested by the novel proteolysis approach to demonstrate its feasibility and performance. The results indicated that AC significantly accelerated in-solution chymotryptic proteolysis and the digestion time was substantially reduced to 5 min. The digests were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with sequence coverages of 46% (BSA) and 90% (Cyt-c) that were much better than those obtained by using 12-h conventional in-solution chymotryptic proteolysis. In addition, AC-assisted chymotryptic proteolysis was employed to digest human serum to demonstrate its suitability to complex protein sample. The present proteolysis strategy is simple and efficient and will find a wide range of applications in proteomic research.
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