Enzyme-encapsulated silica monolayers for rapid functionalization of a gold surface.

We report a simple and rapid method for the deposition of amorphous silica onto a gold surface. The method is based on the ability of lysozyme to mediate the formation of silica nanoparticles. A monolayer of lysozyme is deposited via non-specific binding to gold. The lysozyme then mediates the self-assembled formation of a silica monolayer. The silica formation described herein occurs on a surface plasmon resonance (SPR) gold surface and is characterized by SPR spectroscopy. The silica layer significantly increases the surface area compared to the gold substrate and is directly compatible with a detection system. The maximum surface concentration of lysozyme was found to be a monolayer of 2.6 ng/mm(2) which allowed the deposition of a silica layer of a further 2 ng/mm(2). For additional surface functionalization, the silica was also demonstrated to be a suitable matrix for immobilization of biomolecules. The encapsulation of organophosphate hydrolase (OPH) was demonstrated as a model system. The silica forms at ambient conditions in a reaction that allows the encapsulation of enzymes directly during silica formation. OPH was successfully encapsulated within the silica particles and a detection limit for the substrate, paraoxon, using the surface-encapsulated enzyme was found to be 20 microM.

[1]  Mitchel J. Doktycz,et al.  Comparison of techniques for enzyme immobilization on silicon supports , 1999 .

[2]  B. Simons,et al.  Covalent protein immobilization on glass surfaces: application to alkaline phosphatase. , 2005, Journal of biotechnology.

[3]  ScienceDirect.  Comptes rendus. Palevol , 2002 .

[4]  Hyun-Jong Paik,et al.  Real-time analysis of enzymatic surface-initiated polymerization using surface plasmon resonance (SPR). , 2006, Macromolecular bioscience.

[5]  S. Bram.  Progress in Molecular and Subcellular Biology , 1976 .

[6]  T. Chinowsky,et al.  Quantitative interpretation of the response of surface plasmon resonance sensors to adsorbed films , 1998 .

[7]  Shankar Balasubramanian,et al.  Lytic phage as a specific and selective probe for detection of Staphylococcus aureus--A surface plasmon resonance spectroscopic study. , 2007, Biosensors & bioelectronics.

[8]  Joanna Aizenberg,et al.  Biological glass fibers: correlation between optical and structural properties. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[9]  C. Pace,et al.  Organophosphorus hydrolase is a remarkably stable enzyme that unfolds through a homodimeric intermediate. , 1997, Biochemistry.

[10]  Thomas Laurell,et al.  Silicon wafer integrated enzyme reactors , 1994 .

[11]  S J Tendler,et al.  Surface plasmon resonance analysis of dynamic biological interactions with biomaterials. , 2000, Biomaterials.

[12]  Jeanne E. Pemberton,et al.  Novel Silicon Dioxide Sol-Gel Films for Potential Sensors Applications: A Surface Plasmon Resonance Study , 2001 .

[13]  A. Gast,et al.  Protease adsorption and reaction on an immobilized substrate surface , 2002 .

[14]  Min-Gon Kim,et al.  Enhanced sensitivity of surface plasmon resonance (SPR) immunoassays using a peroxidase-catalyzed precipitation reaction and its application to a protein microarray. , 2005, Journal of immunological methods.

[15]  Haowen Huang,et al.  Label-free reading of microarray-based proteins with high throughput surface plasmon resonance imaging. , 2006, Biosensors & bioelectronics.

[16]  E. G. Vrieling,et al.  Silicon biomineralisation: towards mimicking biogenic silica formation in diatoms. , 2003, Progress in molecular and subcellular biology.

[17]  P. Lopez,et al.  From biogenic to biomimetic silica , 2004 .

[18]  C. Haynes,et al.  Globular proteins at solid/liquid interfaces , 1994 .

[19]  Rajesh R Naik,et al.  Enzyme immobilization in a biomimetic silica support , 2004, Nature Biotechnology.

[20]  Aleksandr L. Simonian,et al.  FET‐Based Biosensors for The Direct Detection of Organophosphate Neurotoxins , 2004 .

[21]  R. Misra,et al.  Biomaterials , 2008 .

[22]  Sabine Szunerits,et al.  Electrochemical investigation of gold/silica thin film interfaces for electrochemical surface plasmon resonance studies , 2006 .

[23]  R. Corn,et al.  Creating advanced multifunctional biosensors with surface enzymatic transformations. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[24]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[25]  K. Lai,et al.  Rational design of organophosphorus hydrolase for altered substrate specificities. , 1999, Chemico-biological interactions.

[26]  장윤희,et al.  Y. , 2003, Industrial and Labor Relations Terms.

[27]  K. Sandhage,et al.  Rapid, room-temperature synthesis of antibacterial bionanocomposites of lysozyme with amorphous silica or titania. , 2006, Small.

[28]  W. Schuhmann,et al.  Reagentless biosensors based on co-entrapment of a soluble redox polymer and an enzyme within an electrochemically deposited polymer film. , 2002, Biosensors & bioelectronics.

[29]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[30]  Joseph S. Schoeniger,et al.  Development of sensors for direct detection of organophosphates.: Part II: sol–gel modified field effect transistor with immobilized organophosphate hydrolase , 1999 .

[31]  John G. Reynolds,et al.  Enzyme Immobilization on Porous Silicon Surfaces , 2004 .

[32]  J. Kleijn,et al.  Adsorption of charged macromolecules at a gold electrode. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[33]  Z. Cui,et al.  The Effect of Solution pH on the Structure of Lysozyme Layers Adsorbed at the Silica−Water Interface Studied by Neutron Reflection , 1998 .

[34]  이현주.  Q. , 2005 .

[35]  Taewook Kang,et al.  Enhancement of surface plasmon resonance (SPR) signals using organic functionalized mesoporous silica on a gold film , 2006 .

[36]  G. Stucky,et al.  Silicatein filaments and subunits from a marine sponge direct the polymerization of silica and silicones in vitro. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Milan Mrksich,et al.  Selective immobilization of proteins to self-assembled monolayers presenting active site-directed capture ligands , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Tsao-Jen Lin,et al.  Determination of organophosphorous pesticides by a novel biosensor based on localized surface plasmon resonance. , 2006, Biosensors & bioelectronics.

[39]  Günter Gauglitz,et al.  Surface plasmon resonance sensors: review , 1999 .

[40]  Jose Melendez,et al.  Detection of Staphylococcus aureus enterotoxin B at femtomolar levels with a miniature integrated two-channel surface plasmon resonance (SPR) sensor. , 2002, Biosensors & bioelectronics.

[41]  F. Young.  Biochemistry , 1955, The Indian Medical Gazette.

[42]  Shiping Fang,et al.  Surface enzyme kinetics for biopolymer microarrays: a combination of Langmuir and Michaelis-Menten concepts. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[43]  N. Kröger,et al.  Polycationic peptides from diatom biosilica that direct silica nanosphere formation. , 1999, Science.