Quantitative and Label-Free Technique for Measuring Protease Activity and Inhibition using a Microfluidic Cantilever Array
- 23 August 2008
- journal article
- letter
- Published by American Chemical Society (ACS) in Nano Letters
- Vol. 8 (9), 2968-2974
- https://doi.org/10.1021/nl8019455
Abstract
We report the use of a SiNx 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 (KM = 0.58 × 10−6 M). Inhibition of surface-immobilized trypsin by soybean trypsin inhibitor (SBTI) was also observed using this system.Keywords
This publication has 41 references indexed in Scilit:
- MEROPS: the peptidase databaseNucleic Acids Research, 2007
- Nano-chemo-mechanical sensor array platform for high-throughput chemical analysisSensors and Actuators B: Chemical, 2006
- Nanomechanical Detection of DNA Melting on Microcantilever SurfacesAnalytical Chemistry, 2006
- Biological Variation of Total Prostate-Specific Antigen: A Survey of Published Estimates and Consequences for Clinical PracticeClinical Chemistry, 2005
- Micromechanical Detection of Proteins Using Aptamer-Based Receptor MoleculesAnalytical Chemistry, 2004
- Protein microarrays and proteomicsNature Genetics, 2002
- Molecular interactions on microarraysNature Genetics, 1999
- Biospecific interaction analysis using biosensor technologyNature, 1993
- Immuno-PCR: Very Sensitive Antigen Detection by Means of Specific Antibody-DNA ConjugatesScience, 1992
- Could domain movements be involved in the mechanism of trypsin‐like serine proteases?FEBS Letters, 1990