title: 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.
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jeong Hoon
authors.initials: JH
authors.lastname: Lee
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Cosgrove
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Lauffenburger
authors.forename: Jongyoon
authors.initials: J
authors.lastname: Han
mesh_terms: D053000:Analytic Sample Preparation Methods; D002457:Cell Extracts / Q000737:chemistry; D045744:Cell Line, Tumor; D002477:Cells / Q000201:enzymology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000378:metabolism*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000378:metabolism*; D000099190:MAP-Kinase-Activated Kinase 2
chemical_list: D002457:Cell Extracts; D017868:Cyclic AMP-Dependent Protein Kinases; D047908:Intracellular Signaling Peptides and Proteins; D017346:Protein Serine-Threonine Kinases; D000099190:MAP-Kinase-Activated Kinase 2