Quantitative and label-free technique for measuring protease activity and inhibition using a microfluidic cantilever array

Nano Lett. 2008 Sep;8(9):2968-74. doi: 10.1021/nl8019455. Epub 2008 Aug 23.


We report the use of a SiN x based gold coated microcantilever array to quantitatively measure the activity and inhibition of a model protease immobilized on its surface. Trypsin was covalently bound to the gold surface of the microcantilever using a synthetic spacer, and the remaining exposed silicon nitride surface was passivated with silanated polyethylene glycol. The nanoscale cantilever motions induced by trypsin during substrate turnover were quantitatively measured using an optical laser-deflection technique. These microcantilever deflections directly correlated with the degree of protease turnover of excess synthetic fibronectin substrate ( K M = 0.58 x 10 (-6) M). Inhibition of surface-immobilized trypsin by soybean trypsin inhibitor (SBTI) was also observed using this system.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Kinetics
  • Microfluidics / methods*
  • Microscopy, Electron, Scanning
  • Peptide Hydrolases / metabolism*
  • Protease Inhibitors / pharmacology*


  • Protease Inhibitors
  • Peptide Hydrolases