Journal
ELECTROPHORESIS
Volume 24, Issue 21, Pages 3679-3688Publisher
WILEY
DOI: 10.1002/elps.200305592
Keywords
cross-linked polymer; electrophoretic separation; miniaturization; poly(dimethylsiloxane); poly(ethylene glycol); ultraviolet grafting
Funding
- NATIONAL CENTER FOR RESEARCH RESOURCES [R01RR014892] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM057015] Funding Source: NIH RePORTER
- NCRR NIH HHS [RR14892] Funding Source: Medline
- NIGMS NIH HHS [GM57015] Funding Source: Medline
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We have developed a strategy using ultraviolet light to polymerize mixed monomer solutions onto the surface of a poly(dimethylsiloxane) (PDMS) microdevice. By including monomers with different chemical properties, electrophoretic separations were optimized for a test set of analytes. The properties of surfaces grafted with a single neutral monomer, a neutral and a negative monomer, or a neutral, negative, and cross-linking monomer were assessed. The highest quality separations were achieved in channels with cross-linked coatings. The separation efficiency for biologically relevant peptides (kinase substrates) on these surfaces was as high as 18 600 theoretical plates in a 2.5 cm channel. The test peptides were fluorescein-AEEEIYGEFEAKKKK, fluorescein-GRPRAATFAEG, fluorescein-GRPRAA(T-PO3)FAEG, fluorescein-DLDVPIP GRFDRRVSVAAE, and fluorescein-DLDVPIPGRFDRRV(S-PO3)VAAE. Separations between two different peptides occurred in as little as 400 ms after injection into the separation channel. The simultaneous separation of five kinase and phosphatase substrates was also demonstrated. By carefully selecting mixtures of monomers with the appropriate properties, it may be possible to tailor the surface of PDMS for a large number of different electrophoretic separations.
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