4.8 Article

Microfluidic Concentration-Enhanced Cellular Kinase Activity Assay

Journal

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 131, Issue 30, Pages 10340-+

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/ja902594f

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Funding

  1. NIH [R01-EB005743, R01-CA119402, P50-GM68672]
  2. DoD Army Institute for Collaborative Biotechnologies

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In this paper, we reported a simple, disposable PDMS micro/nanofluidic preconcentration chip for in vitro concentration-enhanced cell kinase assays. Utilizing the preconcentration (etectrokinetic trapping) directly from cell lysate (1 mM ATP) samples, we could achieve at least a 25-fold increase in reaction velocity and 65-fol.d enhancement in sensitivity. In addition, we shorten the assay time down to less than 10 min, with the sample volume requirements of down to similar to 5 cells. This device could be a generic and powerful tool for diagnostics and systems biology studies at the single-cell level, if property optimized and integrated with the cell culture microdevices.

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