Lysozyme-Mediated Formation of Protein-Silica Nano-Composites for Biosensing Applications (Postprint)

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...

Full description

Saved in:
Bibliographic Details
Main Authors: Ramanathan, Madhumati, Simonian, Aleksandr L, Luckarift, Heather R, Sarsenova, Ainur, Ramanculov, Erlan, Wild, James R, Olsen, Eric V
Format: Report
Language:English
Subjects:
ARRAY BIOSENSOR
Biochemistry
BIODETECTORS
BIOLOGICAL DETECTION
BIOMOLECULES
ENCAPSULATION
ENZYMES
HYDROLASES
LYSOZYMES
Miscellaneous Detection and Detectors
MURAMIDASE
NANOCOMPOSITES
OPH(ORGANOPHOSPHATE HYDROLASE)
ORGANOPHOSPHATES
PARAOXON
PE0602102F
SILICA
SILICIFICATION
SILICON DIOXIDE
WAVEGUIDES
WUAFRLQ140LA62
Online Access:Request full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
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 microM. The encapsulation technique provides a potentially versatile tool with specific application to biosensor development. Document contains color images. Published in Colloids and Surfaces B: Biointerfaces, v73 p58-64, 2009. Sponsored in part by USAF SG approved Clinical Investigation grant no. FDG20060049N and USAF CRADA grant no. 08-272-60MDG-CRADA01, and by the USDA-CSREES under grant no. 2006-34394-16953. Prepared in cooperation with Universal Technology Corporation, Dayton, OH, National Center of Biotechnology of the Republic of Kazakhstan, Astana, Kazakhstan, Texas A&M University, College Station, TX, Air Force Research Laboratory, Tyndall AFB, FL, Clinical Research Facility, 81st Medical Group, Keesler AFB, MS.