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Evaluation of different nonspecific binding blocking agents deposited inside poly(methyl methacrylate) microfluidic flow-cells

Le, N.C.H., Gubala, V., Gandhiraman, R.P., Daniels, S., Williams, D.E. (2011) Evaluation of different nonspecific binding blocking agents deposited inside poly(methyl methacrylate) microfluidic flow-cells. Langmuir, 27 (14). pp. 9043-9051. ISSN 0743-7463. (doi:10.1021/la2011502)

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Abstract

Poly(methyl methacrylate) (PMMA) flow-cells containing microwells were deposited with different nonspecific binding blocking agents, namely, bovine serum albumin (BSA), cationic lipid (DOTAP:DOPE) and diethylene glycol dimethyl ether (DEGDME). Water contact angle (WCA) and atomic force microscope (AFM) measurements were carried out to confirm the successful depositions of BSA, DOTAP, and DEGDME onto the PMMA surfaces. Fluorescent intensity measurements were performed to evaluate the degree of nonspecific adsorption of Cy5-labeled anti-IgG proteins onto plain and oxygen plasma-treated (PT) PMMA flow-cells as well as PMMA flow-cells deposited with different above-mentioned blocking agents. We then employed a label-free detection method called total internal reflection ellipsometry (TIRE) to evaluate the stability of the deposited blocking agents inside the PMMA flow-cells. It was found that, while DOTAP:DOPE was the best agent for blocking the nonspecific adsorption, it could be removed from the PMMA surfaces of the flow-cells upon rinsing with phosphate buffered saline (PBS) and later deposited back onto the Au-coated glass sensing substrate of the TIRE. The removal of the blocking agents from PMMA surfaces and their deposition onto the sensing substrate were further manifested by measuring the kinetics and the amount of adsorbed anti-α-hCG proteins. Overall, the dry DEGDME coating by plasma-enhanced chemical vapor deposition (PECVD) showed very good blocking and excellent stability for subsequent assay inside the microwells. Our results could be useful when one considers what blocking agents should be used for PMMA-based microfluidic immunosensor or biosensor devices by looking at both the blocking efficiency and the stability of the blocking agent. © 2011 American Chemical Society.

Item Type: Article
DOI/Identification number: 10.1021/la2011502
Additional information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Langmuir [Field not mapped to EPrints] C2 - 21648475 [Field not mapped to EPrints] AD - Biomedical Diagnostics Institute (BDI), Dublin City University, Dublin 9, Ireland [Field not mapped to EPrints] AD - National Centre for Plasma Science and Technology (NCPST), Dublin City University, Dublin 9, Ireland [Field not mapped to EPrints] AD - MacDiarmid Institute for Advanced Materials and Nanotechnology, Department of Chemistry, University of Auckland, Auckland-1142, New Zealand [Field not mapped to EPrints] AD - CSIRO Materials Science and Engineering, Highett, VIC 3190, Australia [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled keywords: Atomic force microscopes, Blocking agents, Bovine serum albumins, Cationic lipids, Diethylene glycol, Dimethyl ethers, Fluorescent intensity, Label-free detection method, Microwells, Non-specific binding, Nonspecific adsorption, Phosphate-buffered salines, Total internal reflection ellipsometry, Water contact angle, Adsorption, Atomic force microscopy, Body fluids, Cells, Chemical stability, Chemical vapor deposition, Contact angle, Esters, Ethers, Fuels, Glycols, Immunosensors, Microfluidics, Plasma deposition, Plasma enhanced chemical vapor deposition, Plasma stability, Refractive index, Self assembly, Gold coatings, 1,2 dioleoyl glycero 3 phosphatidyl ethanolamine, 1,2 dioleoyloxy 3 (trimethylammonium)propane, 1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine, 1,2-dioleoyloxy-3-(trimethylammonium)propane, bovine serum albumin, chorionic gonadotropin, diglyme, ether derivative, ethylene glycol derivative, immobilized antibody, monounsaturated fatty acid, phosphatidylethanolamine, poly(methyl methacrylate), quaternary ammonium derivative, adsorption, animal, article, cattle, chemistry, human, immunology, instrumentation, microfluidic analysis, surface property, volatilization, Adsorption, Animals, Antibodies, Immobilized, Cattle, Chorionic Gonadotropin, Ethylene Glycols, Fatty Acids, Monounsaturated, Humans, Methyl Ethers, Microfluidic Analytical Techniques, Phosphatidylethanolamines, Polymethyl Methacrylate, Quaternary Ammonium Compounds, Serum Albumin, Bovine, Surface Properties, Volatilization
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Vladimir Gubala
Date Deposited: 14 Dec 2017 20:09 UTC
Last Modified: 29 May 2019 13:41 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/45228 (The current URI for this page, for reference purposes)
Gubala, V.: https://orcid.org/0000-0001-6301-3632
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