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

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 (A...

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Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1992. - 27(2011), 14 vom: 19. Juli, Seite 9043-51
1. Verfasser: Le, Nam Cao Hoai (VerfasserIn)
Weitere Verfasser: Gubala, Vladimir, Gandhiraman, Ram P, Daniels, Stephen, Williams, David E
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2011
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, Non-U.S. Gov't 1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine Antibodies, Immobilized Chorionic Gonadotropin Ethylene Glycols Fatty Acids, Monounsaturated Methyl Ethers Phosphatidylethanolamines Quaternary Ammonium Compounds mehr... Serum Albumin, Bovine 27432CM55Q Polymethyl Methacrylate 9011-14-7 diglyme M4BH3X0MVZ 1,2-dioleoyloxy-3-(trimethylammonium)propane MR86K0XRQP
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100 1 |a Le, Nam Cao Hoai  |e verfasserin  |4 aut 
245 1 0 |a Evaluation of different nonspecific binding blocking agents deposited inside poly(methyl methacrylate) microfluidic flow-cells 
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520 |a 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 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a Antibodies, Immobilized  |2 NLM 
650 7 |a Chorionic Gonadotropin  |2 NLM 
650 7 |a Ethylene Glycols  |2 NLM 
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650 7 |a Phosphatidylethanolamines  |2 NLM 
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650 7 |a Serum Albumin, Bovine  |2 NLM 
650 7 |a 27432CM55Q  |2 NLM 
650 7 |a Polymethyl Methacrylate  |2 NLM 
650 7 |a 9011-14-7  |2 NLM 
650 7 |a diglyme  |2 NLM 
650 7 |a M4BH3X0MVZ  |2 NLM 
650 7 |a 1,2-dioleoyloxy-3-(trimethylammonium)propane  |2 NLM 
650 7 |a MR86K0XRQP  |2 NLM 
700 1 |a Gubala, Vladimir  |e verfasserin  |4 aut 
700 1 |a Gandhiraman, Ram P  |e verfasserin  |4 aut 
700 1 |a Daniels, Stephen  |e verfasserin  |4 aut 
700 1 |a Williams, David E  |e verfasserin  |4 aut 
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