Trypsin is a serine protease which cleaves peptide bonds after arginine or lysine amino acid residues.  Since the patients who suffer from acute pancreatitis exhibit higher concentrations of trypsin than the healthy people, trypsin is a useful biomarker for pancreatitis.  In this work, a rapid and sensitive method for the detection of trypsin is developed by using a biological alpha-hemolysin nanopore.  Due to its much larger molecular diameter than the narrow pore constriction, trypsin cannot transport through the alpha-hemolysin channel.  Hence, an indirect detection method is developed by using a lysine-containing peptide as the substrate, and analyzing the cleavage product of the substrate after trypsin digestion.  The method is rapid and highly sensitive, with picomolar concentrations of trypsin detected in minutes.  In addition, the salt effect and interference study have also been studied.

Nanopore stochastic sensing of trypsin

248th National Meeting (2014)

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