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Formation of protein molecular imprints within Langmuir monolayers: a quartz crystal microbalance study

Turner, Nicholas; Wright, Bryon E.; Britt, David W. and Hlady, Vladimir (2007). Formation of protein molecular imprints within Langmuir monolayers: a quartz crystal microbalance study. Journal of Colloid and Interface Science, 308(1) 71 -80.

DOI (Digital Object Identifier) Link: http://dx.doi.org/10.1016/j.jcis.2006.12.037
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Abstract

Protein imprinting leading to enhanced rebinding of ferritin to ternary lipid monolayers is demonstrated using a quartz crystal microbalance. Monolayers consisting of cationic dioctadecyldimethylammonium bromide, non-ionic methyl stearate, and poly(ethylene glycol) bearing phospholipids were imprinted with ferritin at the air/water interface of a Langmuir–Blodgett trough and transferred hydrated to hydrophobic substrates for study. This immobilization was shown by fluorescence correlation spectroscopy to significantly hinder any further diffusion of lipids, while rebinding studies demonstrated up to a six-fold increase in ferritin adsorption to imprinted versus control monolayers. A diminished rebinding of ferritin to its imprint was observed through pH reduction to below the protein isoelectric point, demonstrating the electrostatic nature of the interaction. Rebinding to films where imprint pockets remained occupied by the template protein was also minimal. Studies with a smaller acidic protein revealed the importance of the steric influence of poly(ethylene glycol) in forming the protein binding pockets, as albumin-imprinted monolayers showed low binding of ferritin, while ferritin-imprinted monolayers readily accommodated albumin. The controllable structure–function relationship and limitations of this system are discussed with respect to the application of protein imprinting in sensor development as well as fundamental studies of proteins at dynamic interfaces.

Protein imprinting leading to enhanced rebinding of ferritin to ternary lipid monolayers is demonstrated using a quartz crystal microbalance. Monolayers consisting of cationic dioctadecyldimethylammonium bromide, non-ionic methyl stearate, and poly(ethylene glycol) bearing phospholipids were imprinted with ferritin at the air/water interface of a Langmuir–Blodgett trough and transferred hydrated to hydrophobic substrates for study. This immobilization was shown by fluorescence correlation spectroscopy to significantly hinder any further diffusion of lipids, while rebinding studies demonstrated up to a six-fold increase in ferritin adsorption to imprinted versus control monolayers. A diminished rebinding of ferritin to its imprint was observed through pH reduction to below the protein isoelectric point, demonstrating the electrostatic nature of the interaction. Rebinding to films where imprint pockets remained occupied by the template protein was also minimal. Studies with a smaller acidic protein revealed the importance of the steric influence of poly(ethylene glycol) in forming the protein binding pockets, as albumin-imprinted monolayers showed low binding of ferritin, while ferritin-imprinted monolayers readily accommodated albumin. The controllable structure–function relationship and limitations of this system are discussed with respect to the application of protein imprinting in sensor development as well as fundamental studies of proteins at dynamic interfaces.

Item Type: Journal Article
Copyright Holders: 2007 Elsevier
ISSN: 0021-9797
Project Funding Details:
Funded Project NameProject IDFunding Body
Not SetNot SetNational Research Initiative of the USDA Cooperative State Research
Not SetNot SetEducation and Extension Service [2005-35603-15829]
Not SetNot SetNSF-BES Nanoscale Engineering Research [0404262]
Extra Information: Formerly known as Journal of Colloid Science
Academic Unit/Department: Science > Life, Health and Chemical Sciences
Interdisciplinary Research Centre: Centre for Earth, Planetary, Space and Astronomical Research (CEPSAR)
Biomedical Research Network (BRN)
Item ID: 26955
Depositing User: Nicholas Turner
Date Deposited: 26 Jan 2011 18:27
Last Modified: 10 Mar 2014 09:25
URI: http://oro.open.ac.uk/id/eprint/26955
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