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Microfluidic concentration-enhanced cellular kinase activity assay.

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

SUBMITTER: Lee JH

PROVIDER: S-EPMC2739094 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature

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Microfluidic concentration-enhanced cellular kinase activity assay.

Lee Jeong Hoon JH Cosgrove Benjamin D BD Lauffenburger Douglas A DA Han Jongyoon J

Journal of the American Chemical Society 20090801 30

In this paper, we reported a simple, disposable PDMS micro/nanofluidic preconcentration chip for in vitro concentration-enhanced cell kinase assays. Utilizing the preconcentration (electrokinetic trapping) directly from cell lysate (1 mM ATP) samples, we could achieve at least a 25-fold increase in reaction velocity and 65-fold enhancement in sensitivity. In addition, we shorten the assay time down to less than 10 min, with the sample volume requirements of down to approximately 5 cells. This de ...[more]

PMID: 19722608

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