Glucose microfluidic biosensors based on reversible enzyme immobilization on photopatterned stimuli-responsive polymer.

In this paper, we demonstrate a new strategy for replaceable enzymatic microreactor based on a switchable wettability interface of poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm porous polymer monolith (PPM) with 3D macroporous framework is photopolymerized in glass microchip within 30 s. The PNIPAAm PPM not only shows its reversible swelling/shrinking property at the different temperature around the lower critical solution temperature (LCST), but also shows reversible hydrophilicity/hydrophobicity corresponding to its swelling/shrinking status. Based on these properties, a biocompatible and replaceable on-chip enzymatic microreactor has been successfully built by means of the reversible adsorption and release of glucose oxidase (GOx) on the robust and stable matrix. Coupled with a carbon fiber microelectrode as electrochemical detector, the microreactor has been successfully employed for detection of glucose with a linear range from 0.05 to 5 mM. This approach may provide a promising way for high efficient and renewable microreactors that will find wide application in clinical diagnosis, biochemical synthesis/analysis, and proteomic research.

[1]  Hongyuan Chen,et al.  Patterning microbeads inside poly(dimethylsiloxane) microfluidic channels and its application for immobilized microfluidic enzyme reactors , 2006, Electrophoresis.

[2]  Ronald P. Manginell,et al.  Programmed Adsorption and Release of Proteins in a Microfluidic Device , 2003, Science.

[3]  Mwj Menno Prins,et al.  Fluid control in multichannel structures by electrocapillary pressure. , 2001, Science.

[4]  S. Minko,et al.  Stimuli-responsive hydrogel thin films , 2009 .

[5]  Hongyuan Chen,et al.  Glucose microfluidic biosensors based on immobilizing glucose oxidase in poly(dimethylsiloxane) electrophoretic microchips. , 2006, Journal of chromatography. A.

[6]  Ichimura,et al.  Light-driven motion of liquids on a photoresponsive surface , 2000, Science.

[7]  Yan Liu,et al.  In-situ synthesis of poly(dimethylsiloxane)-gold nanoparticles composite films and its application in microfluidic systems. , 2008, Lab on a Chip.

[8]  F. Švec,et al.  Monolithic porous polymer for on-chip solid-phase extraction and preconcentration prepared by photoinitiated in situ polymerization within a microfluidic device. , 2001, Analytical chemistry.

[9]  Hongyuan Chen,et al.  Lab-on-a-chip for analysis of triglycerides based on a replaceable enzyme carrier using magnetic beads. , 2010, The Analyst.

[10]  U. Bilitewski,et al.  Development of monolithic enzymatic reactors in glass microchips for the quantitative determination of enzyme substrates using the example of glucose determination via immobilized glucose oxidase , 2005, Electrophoresis.

[11]  Frantisek Svec,et al.  Flow control valves for analytical microfluidic chips without mechanical parts based on thermally responsive monolithic polymers. , 2003, Analytical chemistry.

[12]  Jing-Juan Xu,et al.  Electrochemical detection method for nonelectroactive and electroactive analytes in microchip electrophoresis. , 2004, Analytical chemistry.

[13]  Jean-Michel Kauffmann,et al.  Preparation, characterization, and application of an enzyme-immobilized magnetic microreactor for flow injection analysis. , 2004, Analytical chemistry.

[14]  J. Rogalski,et al.  Thermoresponsive poly(N-isopropylacrylamide) gel for immobilization of laccase on indium tin oxide electrodes. , 2009, The journal of physical chemistry. B.

[15]  Frantisek Svec,et al.  Monolithic valves for microfluidic chips based on thermoresponsive polymer gels , 2003, Electrophoresis.

[16]  Shan‐Yang Lin,et al.  Thermal micro ATR/FT-IR spectroscopic system for quantitative study of the molecular structure of poly(N-isopropylacrylamide) in water , 1999 .

[17]  Hideaki Hisamoto,et al.  Integration of valving and sensing on a capillary-assembled microchip. , 2005, Analytical chemistry.

[18]  Patrick S. Stayton,et al.  Conjugates of stimuli-responsive polymers and proteins , 2007 .

[19]  Lei Jiang,et al.  Reversible switching between superhydrophilicity and superhydrophobicity. , 2004, Angewandte Chemie.

[20]  Amy E Herr,et al.  Automated microfluidic protein immunoblotting , 2010, Nature Protocols.

[21]  Amy E Herr,et al.  Photopatterned materials in bioanalytical microfluidic technology , 2011, Journal of micromechanics and microengineering : structures, devices, and systems.

[22]  Masayuki Yamato,et al.  Ultrathin poly(N-isopropylacrylamide) grafted layer on polystyrene surfaces for cell adhesion/detachment control. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[23]  L. Jiang,et al.  Multiresponsive Surfaces Change Between Superhydrophilicity and Superhydrophobicity , 2007 .

[24]  S. Soper,et al.  Thermoswitchable electrokinetic ion-enrichment/elution based on a poly(N-isopropylacrylamide) hydrogel plug in a microchannel. , 2010, Analytical Chemistry.

[25]  Robin H. Liu,et al.  Functional hydrogel structures for autonomous flow control inside microfluidic channels , 2000, Nature.

[26]  Zeng-Qiang Wu,et al.  Exploring the temperature-dependent kinetics and thermodynamics of immobilized glucose oxidase in microchip , 2012 .

[27]  G. Whitesides The origins and the future of microfluidics , 2006, Nature.

[28]  V. Choudhary,et al.  Synthesis and characterization of poly(N‐isopropylacrylamide) films by photopolymerization , 2006 .

[29]  Qiaohong He,et al.  In-channel modification of biosensor electrodes integrated on a polycarbonate microfluidic chip for micro flow-injection amperometric determination of glucose , 2010 .

[30]  H. Ju,et al.  Open Tubular Microreactor with Enzyme Functionalized Microfluidic Channel for Amperometric Detection of Glucose , 2012 .

[31]  Hideaki Maeda,et al.  Microchannel enzyme reactors and their applications for processing. , 2006, Trends in biotechnology.

[32]  Zhiming Li,et al.  On-chip integrated multi-thermo-actuated microvalves of poly(N-isopropylacrylamide) for microflow injection analysis. , 2010, Analytica chimica acta.

[33]  Jing-Juan Xu,et al.  Off-line form of the Michaelis-Menten equation for studying the reaction kinetics in a polymer microchip integrated with enzyme microreactor. , 2006, Lab on a chip.

[34]  Zeng-Qiang Wu,et al.  Real-time monitoring of mass-transport-related enzymatic reaction kinetics in a nanochannel-array reactor. , 2010, Chemistry.

[35]  Jing‐Juan Xu,et al.  Gold nanoparticles-coated magnetic microspheres as affinity matrix for detection of hemoglobin A1c in blood by microfluidic immunoassay. , 2011, Biosensors & bioelectronics.

[36]  Patrick F. Kiser,et al.  A synthetic mimic of the secretory granule for drug delivery , 1998, Nature.

[37]  P. C. Rieke,et al.  Reversible Surface Properties of Glass Plate and Capillary Tube Grafted by Photopolymerization of N-Isopropylacrylamide , 1998 .

[38]  Frantisek Svec,et al.  Enzymatic microreactor-on-a-chip: protein mapping using trypsin immobilized on porous polymer monoliths molded in channels of microfluidic devices. , 2002, Analytical chemistry.

[39]  T. Russell,et al.  Surface-Responsive Materials , 2002, Science.

[40]  Jin Sheng,et al.  Fabrication of tunable microreactor with enzyme modified magnetic nanoparticles for microfluidic electrochemical detection of glucose. , 2012, Analytica chimica acta.

[41]  Zhao-Lun Fang,et al.  Bonding of glass microfluidic chips at room temperatures. , 2004, Analytical chemistry.

[42]  Ying Liu,et al.  A smart surface in a microfluidic chip for controlled protein separation. , 2007, Chemistry.