Microfluidic bead-based sensing platform for monitoring kinase activity

• Published in: Biosensors & bioelectronics 2014-07, Vol.57, p.1-9
• Main Authors: Lee, Seung Hwan, Rhee, Hyun-Woo, van Noort, Danny, Lee, Hong Jai, Park, Hee Ho, Shin, Ik-Soo, Hong, Jong-In, Park, Tai Hyun
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biosensing Techniques - instrumentation; Biotechnology; Chemosensor; Enzyme Assays - instrumentation; Equipment Design; Fundamental and applied biological sciences. Psychology; HeLa Cells; Humans; Kinase assay; Microfluidic Analytical Techniques - instrumentation; Microfluidic device; Peptides - metabolism; Phosphorylation; Protein kinase activity; Protein Kinases - metabolism
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2014.01.039

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. •A novel microfluidic system was developed to monitor kinase activity using a bead-based turn-on chemosensor.•The integrated DPA beads discriminated between phosphorylated and non-phosphorylated peptides based on their fluorescence signal.•The activities of various kinases and the inhibitory effect of specific chemicals on the kinases were monitored.•The repeatability of the kinase activity monitoring system was confirmed for multiple assays.