Lysozyme-mediated formation of protein–silica nano-composites for biosensing applications

Lysozyme-mediated formation of protein–silica nano-composites for biosensing applications

We demonstrate a rapid method for enzyme immobilization directly on a waveguide surface by encapsulation in a silica matrix. Organophosphate hydrolase (OPH), an enzyme that catalytically hydrolyzes organophosphates, was used as a model enzyme to demonstrate the utility of lysozyme-mediated silica fo...

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Bibliographic Details
Published in:Colloids and surfaces, B, Biointerfaces Vol. 73; no. 1; pp. 58 - 64
Main Authors: Ramanathan, Madhumati, Luckarift, Heather R., Sarsenova, Ainur, Wild, James R., Ramanculov, Erlan K., Olsen, Eric V., Simonian, Aleksandr L.
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01-10-2009
Subjects:
Animals
Array biosensor
Biosensing Techniques - instrumentation
Biosensing Techniques - methods
Chickens
Enzymes, Immobilized - metabolism
Eyeglasses
Female
Hydrolysis
Lysozyme
Microscopy, Electron, Scanning
Molecular Structure
Muramidase - metabolism
Nanoparticles - chemistry
Nanoparticles - ultrastructure
Nitrophenols - chemistry
Nitrophenols - metabolism
OPH
Paraoxon
Paraoxon - chemistry
Paraoxon - metabolism
Phosphoric Monoester Hydrolases - chemistry
Phosphoric Monoester Hydrolases - metabolism
Silica
Silicification
Silicon Dioxide - chemistry
Array biosensor
Silica
OPH
Paraoxon
Lysozyme
Silicification
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Summary:We demonstrate a rapid method for enzyme immobilization directly on a waveguide surface by encapsulation in a silica matrix. Organophosphate hydrolase (OPH), an enzyme that catalytically hydrolyzes organophosphates, was used as a model enzyme to demonstrate the utility of lysozyme-mediated silica formation for enzyme stabilization. Silica morphology and the efficiency of OPH encapsulation were directly influenced by the precursor choice used in silica formation. Covalent attachment of the lysozyme template directly to the waveguide surface provided a stable basis for silica formation and significantly increased the surface area for OPH encapsulation. OPH conjugated to a pH-responsive fluorophore was encapsulated in silica and patterned to a waveguide surface to demonstrate the immobilization strategy for the development of an organophosphate array biodetector. Silica-encapsulated OPH retained its catalytic activity for nearly 60 days with a detection limit of paraoxon of ∼35 μM. The encapsulation technique provides a potentially versatile tool with specific application to biosensor development.
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ISSN:0927-7765
1873-4367
DOI:10.1016/j.colsurfb.2009.04.024