title: Quantitative and label-free technique for measuring protease activity and inhibition using a microfluidic cantilever array.
abstract: 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.
authors: Department of Mechanical Engineering, University of California, Berkeley, California 94720, USA.
authors: Digvijay A
authors: DA
authors: Raorane
authors: Mark D
authors: MD
authors: Lim
authors: Fanqing Frank
authors: FF
authors: Chen
authors: Charles S
authors: CS
authors: Craik
authors: Arun
authors: A
authors: Majumdar
mesh_terms: D007700:Kinetics; D044085:Microfluidics / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology*
chemical_list: D011480:Protease Inhibitors; D010447:Peptide Hydrolases