Application of plasma-polymerized films for isoelectric focusing of proteins in a capillary electrophoresis chip

The first use of plasma polymerization technique to modify the surface of a glass chip for capillary isoelectric focusing (cIEF) of different proteins is reported. The electrophoresis separation channel was machined in Tempax glass chips with length 70 mm, 300 microm width and 100 microm depth. Acet...

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Published inAnalyst (London) Vol. 128; no. 3; pp. 237 - 244
Main Authors TSAI, Shuo-Wen, LOUGHRAN, Michael, HIRATSUKA, Atsunori, YANO, Kazuyoshi, KARUBE, Isao
Format Journal Article
LanguageEnglish
Published Cambridge Royal Society of Chemistry 01.03.2003
Subjects
Analytical chemistry
Animals
Chemistry
Chromatographic methods and physical methods associated with chromatography
Electrophoresis, Capillary - instrumentation
Exact sciences and technology
Glass
Isoelectric Focusing - instrumentation
Isoelectric Focusing - methods
Other chromatographic methods
Polymers
Proteins - analysis
Capillary electrophoresis
Protein
Isoelectric focusing
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Summary:The first use of plasma polymerization technique to modify the surface of a glass chip for capillary isoelectric focusing (cIEF) of different proteins is reported. The electrophoresis separation channel was machined in Tempax glass chips with length 70 mm, 300 microm width and 100 microm depth. Acetonitrile and hexamethyldisiloxane monomers were used for plasma polymerization. In each case 100 nm plasma polymer films were coated onto the chip surface to reduce protein wall adsorption and minimize the electroosmotic flow. Applied voltages of 1000 V, 2000 V and 3000 V were used to separate mixtures of cytochrome c (pI 9.6), hemoglobin (pI 7.0) and phycocyanin (pI 4.65). Reproducible isoelectric focusing of each pI marker protein was observed in different coated capillaries at increasing concentration 2.22-5 microg microL(-1). Modification of the glass capillary with hydrophobic HMDS plasma polymerized films enabled rapid cIEF within 3 min. The separation efficiency of cytochrome c and phycocyanin in both acrylamide and HMDS coated capillaries corresponded to a plate number of 19600 which compares favourably with capillary electrophoresis of neurotransmitters with amperometric detection.
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ISSN:0003-2654
1364-5528
DOI:10.1039/b207871f