title: Microfluidic bead-based sensing platform for monitoring kinase activity.
abstract: Protein kinases control cellular functions by regulating protein phosphorylation. Monitoring protein kinase activity is essential for medical diagnosis and drug screening. Here, we present a novel microfluidic device for performing simple and versatile protein kinase assays, which utilizes a microbead-based chemosensor. An automatic mix-and-measure technique was achieved using integrated pneumatic valves. After mixing each reagent for the kinase assay, the mixture was transferred to the sensing chamber. Then, phosphorylated and fluorescence-labeled peptides were captured and detected by the chemosensor. A fluorescence signal was observed depending on the presence of the kinase. Furthermore, activities of various kinases in the cell lysate and the inhibitory effect of specific chemicals on the kinases were monitored. These results indicate that chemosensor-based microfluidic chips can be developed as a versatile kinase assay system.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-744, Republic of Korea.
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
authors.affiliation: School of Nano-Bioscience and Chemical Engineering, Ulsan National Institute of Science and Technology, 100 Banyeon-ri, Ulju-gun, Ulsan 689-798, Republic of Korea.
authors.forename: Hyun-Woo
authors.initials: HW
authors.lastname: Rhee
authors.affiliation: Department of Chemistry and Nano Sciences, Ewha Womans University, 11-1 Daehyun-dong, Seodaemun-gu, Seoul 120-750, Republic of Korea.
authors.forename: Danny
authors.initials: D
authors.lastname: van Noort
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-744, Republic of Korea.
authors.forename: Hong Jai
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-744, Republic of Korea.
authors.forename: Hee Ho
authors.initials: HH
authors.lastname: Park
authors.affiliation: Department of Chemistry, Soongsil University, 369 Sangdo-ro, Dongjak-gu, Seoul 156-743, Republic of Korea.
authors.forename: Ik-Soo
authors.initials: IS
authors.lastname: Shin
authors.affiliation: Department of Chemistry, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-747, Republic of Korea. Electronic address: jihong@snu.ac.kr.
authors.forename: Jong-In
authors.initials: JI
authors.lastname: Hong
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-744, Republic of Korea; Advanced Institutes of Convergence Technology, Suwon, Gyeonggi-do 443-270, Republic of Korea. Electronic address: thpark@snu.ac.kr.
authors.forename: Tai Hyun
authors.initials: TH
authors.lastname: Park
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D057075:Enzyme Assays / Q000295:instrumentation; D004867:Equipment Design; D006367:HeLa Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D010455:Peptides; D011494:Protein Kinases
keywords: Chemosensor; Kinase assay; Microfluidic device; Protein kinase activity