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Probing fundamental film parameters of immobilized enzymes—Towards enhanced biosensor performance. Part I—QCM-D mass and rheological measurements

Paper ID Volume ID Publish Year Pages File Format Full-Text
17398 42666 2011 7 PDF Available
Title
Probing fundamental film parameters of immobilized enzymes—Towards enhanced biosensor performance. Part I—QCM-D mass and rheological measurements
Abstract

Enzyme immobilization is an ever-growing research-area for both analytical and industrial applications. Of critical importance in this area are the effects of immobilization procedures upon the functionality of the immobilized biomolecules. Both beneficial and detrimental effects can be conferred through the selection and tuning of the immobilization procedure. Quartz-crystal microbalance with dissipation (QCM-D) has been previously used to great effect in tracking alterations to thin films of biomolecules immobilized onto quartz transducers.In this study, we investigate the ability of QCM-D to track and monitor film parameters of a monolayer of laccase immobilized on a series of self-assembled monolayers (SAMs), differing in lateral density of binding residues on the SAM and height of the SAM from the quartz surface. Both mass gains and rheological parameters for these varying surfaces were measured and trends later compared to the apparent enzyme kinetics of the immobilized laccase films, assessed electroanalytically (Paper II in this two part study). For covalent attachment of proteins, both shear and viscosity were increased relative to physically adsorbed proteins. An increase in lateral density of protein-binding surface of the SAM components was shown to increase the shear/viscosity of the resultant film while an increase in distance from the electrode (through incorporation of lysine linkers) was shown to decrease the shear/viscosity while simultaneously increasing the wet mass gain of the films. Shear and viscosity may be indicative of both enzyme denaturation and increased lateral protein packing within the film structure hence it is assumed that less distortion occurs with the inclusion of linkers which allow for more optimal protein immobilization.

Keywords
QCM-D, quartz-crystal microbalance with dissipation; SAM, self-assembled monolayer; SLB, succinic-lactic acid buffer pH 4.5; V.I., viscoelasticity index; f, frequency; D, dissipation; SPR, surface plasmon resonance; AFM, atomic force microscopyImmobilizat
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Probing fundamental film parameters of immobilized enzymes—Towards enhanced biosensor performance. Part I—QCM-D mass and rheological measurements
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 49, Issue 2, 10 July 2011, Pages 146–152
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us