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Separations in poly(dimethylsiloxane) microchips coated with supported bilayer membranes.

ABSTRACT: Hybrid microchannels composed of poly(dimethylsiloxane) and glass were coated with supported bilayer membranes (SBMs) by the process of vesicle fusion. The electroosmotic mobility (mu(eo)) of zwitterionic, positively charged, and negatively charged phospholipid membranes was measured over a 4 h time to evaluate the stability of the coatings in an electric field. Coated microchips with a simple cross design were used to separate the fluorescent dyes fluorescein and Oregon Green. Migration time reproducibility was better than 5% RSD over 70 min of continuous separations. Separation of Oregon Green and fluorescein in channels coated with zwitterionic phosphatidylcholine (PC) membranes yielded efficiencies of 611,000 and 499,000 plates/m and a resolution of 2.4 within 2 s. Both zwitterionic and negatively charged membranes were used to separate peptide substrates from their phosphorylated analogues with efficiencies of 200,000-400,000 plates/m. Notably, separations of fluorescently labeled ABL substrate peptide from its phosphorylated counterpart were achieved using a high-salt physiological buffer with near-baseline resolution in 10 s. PC-coated devices were used to successfully separate enhanced green fluorescent protein (eGFP) from a fusion protein (eGFP-Crakl) with an efficiency of 358,000 and 278,000 plates/m respectively in less than 12 s. These SBM-based coatings may enable the separation of a broad range of analytes and may be ideal in biological applications for microfluidics.

SUBMITTER: Phillips KS

PROVIDER: S-EPMC2735572 | biostudies-literature | 2008 Dec

REPOSITORIES: biostudies-literature

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Separations in poly(dimethylsiloxane) microchips coated with supported bilayer membranes.

Phillips K Scott KS Kottegoda Sumith S Kang Kyung Mo KM Sims Christopher E CE Allbritton Nancy L NL

Analytical chemistry 20081201 24

Hybrid microchannels composed of poly(dimethylsiloxane) and glass were coated with supported bilayer membranes (SBMs) by the process of vesicle fusion. The electroosmotic mobility (mu(eo)) of zwitterionic, positively charged, and negatively charged phospholipid membranes was measured over a 4 h time to evaluate the stability of the coatings in an electric field. Coated microchips with a simple cross design were used to separate the fluorescent dyes fluorescein and Oregon Green. Migration time re ...[more]

PMID: 19006406

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Project description:Protein-reinforced supported bilayer membranes (rSBMs) composed of phosphatidylcholine (PC), biotin-PE and Neutravidin were used to coat hybrid microchips composed of polydimethylsiloxane (PDMS) and glass. Since the coatings required a freshly oxidized, hydrophilic substrate, a novel method to rapidly connect reservoirs using plasma oxidation was first developed and found to support up to 5.2 N cm(-2) (1.5 N) pull-off force. rSBMs were then assembled in the oxidized hydrophilic channels. The electroosmotic mobility (mu(eo)) of rSBM-coated channels was measured over a 3 h time to evaluate the stability of the coatings for microchip electrophoresis. rSBM-coated microchips with a simple cross-design had excellent properties for microchip separations, yielding efficiencies of up to 700,000 plates m(-1) for fluorescent dyes and peptides. The separation performance of rSBM and PC-coated channels was evaluated after repeatedly drying and rehydrating the channels. The separation efficiency of fluorescein on PC-coated devices decreased by 40% after one dehydration cycle and nearly 75% after 3 cycles. In contrast for rSBM-coated devices there was no significant change in the fluorescein efficiency until the third cycle (10% decreased efficiency). rSBM-coated channels were also markedly more stable when placed in a dehydrated state during long-term storage compared to PC-coated channels, and showed reduced chip failure and no reduction in performance for up to one month of dehydrated storage. Finally, rSBM-coated devices were used to perform single-cell cytometry. Microchips that had been dehydrated, stored two weeks, and rehydrated prior to use demonstrated similar performance to newly coated devices for the separation of fluorescein and carboxyfluorescein from single cells. Thus rSBM-coated devices were rugged withstanding electric fields, prolonged storage under dehydrated conditions, and biofouling by cellular constituents while maintaining excellent separation performance.

Dataset Information

Separations in poly(dimethylsiloxane) microchips coated with supported bilayer membranes.

Publications

Separations in poly(dimethylsiloxane) microchips coated with supported bilayer membranes.

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