Format

Send to

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

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

Author information

1
Department of Mechanical Engineering, University of California, Berkeley, California 94720, USA.

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.

PMID:
18720973
PMCID:
PMC2663003
DOI:
10.1021/nl8019455
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for American Chemical Society Icon for PubMed Central
Loading ...
Support Center