Construction and Characterization of Porous SiO2/Hydrogel Hybrids as Optical Biosensors for Rapid Detection of Bacteria

The use of a new class of hybrid nanomaterials as label-free optical biosensors for bacteria detection (E. coli K12 as a model system) is demonstrated. The hybrids combine a porous SiO 2 (PSiO 2 ) optical nanostructure (a Fabry―Perot thin film) used as the optical transducer element and a hydrogel. The hydrogel, polyacrylamide, is synthesized in situ within the nanostructure inorganic host and conjugated with specific monoclonal antibodies (IgGs) to provide the active component of the biosensor. The immobilization of the IgGs onto the hydrogel via a biotin-streptavidin system is confirmed by fluorescent labeling experiments and reflective interferometric Fourier transform spectroscopy (RIFTS). Additionally, the immobilized IgGs maintain their immunoactivity and specificity when attached to the sensor surface. Exposure of these modified-hybrids to the target bacteria results in "direct cell capture" onto the biosensor surface. These specific binding events induce predictable changes in the thin-film optical interference spectrum of the hybrid. Preliminary studies demonstrate the applicability of these biosensors for the detection of low bacterial concentrations in the range of 10 3 ―10 5 cell mL ―1 within minutes.

[1]  Michael J Sailor,et al.  Covalent crosslinking of 1-D photonic crystals of microporous Si by hydrosilylation and ring-opening metathesis polymerization. , 2003, Chemical communications.

[2]  D. Szarowski,et al.  Biological functionalization and surface micropatterning of polyacrylamide hydrogels. , 2006, Biomaterials.

[3]  Kristopher A Kilian,et al.  The importance of surface chemistry in mesoporous materials: lessons from porous silicon biosensors. , 2009, Chemical communications.

[4]  Michael J. Sailor,et al.  A Porous Silicon Optical Biosensor: Detection of Reversible Binding of IgG to a Protein A-Modified Surface , 1999 .

[5]  Michael J. Sailor,et al.  Polymer Replicas of Photonic Porous Silicon for Sensing and Drug Delivery Applications , 2003, Science.

[6]  Benjamin L Miller,et al.  Cross-correlation of optical microcavity biosensor response with immobilized enzyme activity. Insights into biosensor sensitivity. , 2005, Analytical chemistry.

[7]  J. Buriak,et al.  Chemical and Biological Applications of Porous Silicon Technology , 2000 .

[8]  Kristopher A Kilian,et al.  Smart tissue culture: in situ monitoring of the activity of protease enzymes secreted from live cells using nanostructured photonic crystals. , 2009, Nano letters.

[9]  G. Demirel,et al.  Construction of a novel multilayer system and its use for oriented immobilization of immunoglobulin G , 2007 .

[10]  David S. Wishart,et al.  The CyberCell Database (CCDB): a comprehensive, self-updating, relational database to coordinate and facilitate in silico modeling of Escherichia coli , 2004, Nucleic Acids Res..

[11]  J. Devoisselle,et al.  Confinement of Thermoresponsive Hydrogels in Nanostructured Porous Silicon Dioxide Templates , 2007 .

[12]  Nanostructured silicon-based biosensors for the selective identification of analytes of social interest , 2006 .

[13]  Michael J. Sailor,et al.  Using a porous silicon photonic crystal for bacterial cell‐based biosensing , 2007 .

[14]  Michael J. Sailor,et al.  Protein‐Coated Porous‐Silicon Photonic Crystals for Amplified Optical Detection of Protease Activity , 2006 .

[15]  Michael J Sailor,et al.  Reflective interferometric fourier transform spectroscopy: a self-compensating label-free immunosensor using double-layers of porous SiO2. , 2006, Journal of the American Chemical Society.

[16]  Michael J Sailor,et al.  Biosensing using porous silicon double-layer interferometers: reflective interferometric Fourier transform spectroscopy. , 2005, Journal of the American Chemical Society.

[17]  Philippe M. Fauchet,et al.  Hydrogel‐Supported Optical‐Microcavity Sensors , 2005 .

[18]  J. Hoheisel,et al.  Solid supports for microarray immunoassays , 2003, Journal of molecular recognition : JMR.

[19]  P. Skottrup,et al.  Towards on-site pathogen detection using antibody-based sensors. , 2008, Biosensors & bioelectronics.

[20]  E. Alocilja,et al.  A high density microelectrode array biosensor for detection of E. coli O157:H7. , 2005, Biosensors & bioelectronics.

[21]  E. Alocilja,et al.  Characterization of Nanoporous Silicon-Based DNA Biosensor for the Detection of Salmonella Enteritidis , 2008, IEEE Sensors Journal.

[22]  J. Hoheisel,et al.  Antibody microarrays: An evaluation of production parameters , 2003, Proteomics.

[23]  X. G. Zhang.  Morphology and Formation Mechanisms of Porous Silicon , 2004 .

[24]  Micha E. Spira,et al.  Neurons culturing and biophotonic sensing using porous silicon , 2004 .

[25]  P. Fauchet,et al.  Identification of Gram negative bacteria using nanoscale silicon microcavities. , 2001, Journal of the American Chemical Society.

[26]  Huimin Ouyang,et al.  Label-free quantitative detection of protein using macroporous silicon photonic bandgap biosensors. , 2007, Analytical chemistry.

[27]  E. Segal,et al.  Preparation and Characterization of a pH‐ and Thermally Responsive Poly(N‐isopropylacrylamide‐co‐acrylic acid)/Porous SiO2 Hybrid , 2010, Advanced functional materials.

[28]  Michael J. Sailor,et al.  Chitosan Hydrogel‐Capped Porous SiO2 as a pH Responsive Nano‐Valve for Triggered Release of Insulin , 2009 .

[29]  S. Ossicini,et al.  Porous silicon: a quantum sponge structure for silicon based optoelectronics , 2000 .

[30]  Michael J Sailor,et al.  "Smart dust": nanostructured devices in a grain of sand. , 2005, Chemical communications.

[31]  Michael J Sailor,et al.  The smart Petri dish: a nanostructured photonic crystal for real-time monitoring of living cells. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[32]  Nicolas H Voelcker,et al.  Porous silicon biosensors on the advance. , 2009, Trends in biotechnology.

[33]  Sara D. Alvarez,et al.  Porous SiO2 interferometric biosensor for quantitative determination of protein interactions: binding of protein A to immunoglobulins derived from different species. , 2007, Analytical chemistry.

[34]  J. Pyun,et al.  Immunosensor with a controlled orientation of antibodies by using NeutrAvidin-protein A complex at immunoaffinity layer. , 2006, Journal of biotechnology.

[35]  Andreas Janshoff,et al.  Macroporous p-Type Silicon Fabry−Perot Layers. Fabrication, Characterization, and Applications in Biosensing , 1998 .