Photonic Crystal Kinase Biosensor

We have developed a novel biosensor for kinases that is based on a kinase-responsive polymer hydrogel, which enables label-free screening of kinase activity via changes in optical properties. The hydrogel is specifically designed to swell reversibly upon phosphorylation of a target peptide, triggering a change in optical diffraction from a crystalline colloidal array of particles impregnated into the hydrogel. Diffraction measurements, and charge staining, confirmed the responsive nature of the hydrogel. Moreover, the change in diffraction of the hydrogel upon treatment with kinase exhibited a time- and dose-dependent response. A theoretical model for ionic polymer networks describes the observed optical response well and can be used to quantify the extent of phosphorylation.

[1]  J. Anderson,et al.  Partitioning and diffusion of proteins and linear polymers in polyacrylamide gels. , 1996, Biophysical journal.

[2]  Justin T. Baca,et al.  Progress in developing polymerized crystalline colloidal array sensors for point-of-care detection of myocardial ischemia. , 2008, The Analyst.

[3]  P. Hajduk,et al.  Navigating the kinome. , 2011, Nature chemical biology.

[4]  S. Asher,et al.  Polymerized colloidal crystal hydrogel films as intelligent chemical sensing materials , 1997, Nature.

[5]  High-throughput kinase assay based on surface plasmon resonance. , 2010, Methods in molecular biology.

[6]  Youyong Li,et al.  Discovery and optimization of triazine derivatives as ROCK1 inhibitors: molecular docking, molecular dynamics simulations and free energy calculations. , 2013, Molecular bioSystems.

[7]  Reese,et al.  Synthesis of Highly Charged, Monodisperse Polystyrene Colloidal Particles for the Fabrication of Photonic Crystals. , 2000, Journal of colloid and interface science.

[8]  T. Niidome,et al.  Measurement of Homogeneous Kinase Activity for Cell Lysates Based on the Aggregation of Gold Nanoparticles , 2007, Chembiochem : a European journal of chemical biology.

[9]  Dario R Alessi,et al.  Kinase drug discovery--what's next in the field? , 2013, ACS chemical biology.

[10]  A. Lochner,et al.  The many faces of H89: a review. , 2006, Cardiovascular drug reviews.

[11]  W. DeGrado,et al.  Nature-inspired design of motif-specific antibody scaffolds , 2013, Nature Biotechnology.

[12]  M. Fischer,et al.  Surface plasmon resonance : methods and protocols , 2010 .

[13]  Y. Ikada,et al.  Sorption of low-molecular-weight anions into thin polycation layers grafted onto a film , 1993 .

[14]  Stefan Knapp,et al.  Kinase inhibitor selectivity profiling using differential scanning fluorimetry. , 2012, Methods in molecular biology.

[15]  Kyong‐Cheol Ko,et al.  A facile method for detecting calcium/calmodulin-dependent protein kinase using radio phosphorylation of a GBP-fused enzyme substrate in a lab-on-a-chip , 2013 .

[16]  S. Yao,et al.  Label-free fluorescent detection of protein kinase activity based on the aggregation behavior of unmodified quantum dots. , 2011, Analytical chemistry.

[17]  Y. Ikada,et al.  Mechanism of amide formation by carbodiimide for bioconjugation in aqueous media. , 1995, Bioconjugate chemistry.

[18]  Takeshi Mori,et al.  Optimal surface chemistry for peptide immobilization in on-chip phosphorylation analysis. , 2008, Analytical chemistry.

[19]  P. Flory.  Principles of polymer chemistry , 1953 .

[20]  T. Niidome,et al.  A simple set-and-mix assay for screening of protein kinase inhibitors in cell lysates. , 2011, Analytical biochemistry.

[21]  Jiali Gao,et al.  Dynamically committed, uncommitted, and quenched states encoded in protein kinase A revealed by NMR spectroscopy , 2011, Proceedings of the National Academy of Sciences.

[22]  Randall W King,et al.  High-throughput kinase profiling: a more efficient approach toward the discovery of new kinase inhibitors. , 2011, Chemistry & biology.

[23]  S. Asher,et al.  Acetylcholinesterase-based organophosphate nerve agent sensing photonic crystal. , 2005, Analytical chemistry.

[24]  T. Hunter,et al.  Signaling—2000 and Beyond , 2000, Cell.

[25]  C. Prével,et al.  Fluorescent biosensors for high throughput screening of protein kinase inhibitors , 2014, Biotechnology journal.

[26]  Masaki Inagaki,et al.  Detection of vimentin serine phosphorylation by multicolor Quenchbodies. , 2013, Biosensors & bioelectronics.

[27]  T. Jana,et al.  Photonic crystal hydrogel material for the sensing of toxic mercury ions (Hg2+) in water , 2011 .

[28]  Lei Liu,et al.  Self-Assembly Motif for Creating Submicron Periodic Materials. Polymerized Crystalline Colloidal Arrays , 1994 .

[29]  G. Drewes,et al.  Mass spectrometry approaches to monitor protein-drug interactions. , 2012, Methods.

[30]  Anjal C. Sharma,et al.  A general photonic crystal sensing motif: creatinine in bodily fluids. , 2004, Journal of the American Chemical Society.

[31]  Makoto Hashizume,et al.  Gold nanoparticle-based colorimetric assay for cancer diagnosis. , 2010, Biosensors & bioelectronics.

[32]  T. Jana,et al.  Tuning the particle size and charge density of the crosslinked polystyrene particles , 2008 .

[33]  Shouzhuo Yao,et al.  Aptameric peptide for one-step detection of protein kinase. , 2012, Analytical chemistry.

[34]  A. Ravve,et al.  Principles of Polymer Chemistry , 1995 .