Separations of proteins and proteolytic digests of proteins by capillary electrophoresis on superox‐coated open tubular columns

Zhao, Zhiwei; Malik, Abdul; Lee, Milton L.

doi:10.1002/mcs.1220040508

Cited by 13 publications

(7 citation statements)

References 31 publications

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“…The best results were obtained using PEG or PEG-like polymers. [36][37][38][39][40] In the present work, we describe a technique using ATRP to graft PEG on the surface of PMMA µCE devices for electrophoretic separations of proteins and peptides.…”

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confidence: 99%

Surface-Modified Poly(methyl methacrylate) Capillary Electrophoresis Microchips for Protein and Peptide Analysis

et al. 2004

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Polymeric materials have emerged as appealing alternatives to conventional inorganic substrates for the fabrication of microscale analytical systems; however, native polymeric surfaces typically require covalent modification to ensure optimum biocompatibility. 2-Bromoisobutyryl bromide was immobilized on poly(methyl methacrylate) (PMMA) substrates activated using an oxygen plasma. Atom-transfer radical polymerization was then performed to graft poly(ethylene glycol) (PEG) on the PMMA surface. PMMA microcapillary electrophoresis (μCE) devices made with the covalently modified surfaces exhibited substantially reduced electroosmotic flow and nonspecific adsorption of proteins on microchannel surfaces. Experiments using fluorescein isothiocyanate-conjugated bovine serum albumin indicated that both column efficiency and migration time reproducibility were 1 order of magnitude better with derivatized compared to untreated PMMA μCE chips. Fast, reproducible, and efficient separations of proteins and peptides were demonstrated using the PEG-grafted PMMA μCE chips. All analyses were completed in less than 60 s, and separation efficiencies as high as 5.2 × 104 plates for a 3.5-cm-long separation channel were obtained. These results demonstrate the general applicability of surface-grafted PMMA microdevices for a broad range of protein analyses.

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Surface-Modified Poly(methyl methacrylate) Capillary Electrophoresis Microchips for Protein and Peptide Analysis

et al. 2004

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“…Our results show that the use of polymer-coated capillary columns could eventually become a fast, simple, and efficient alternative for the separation and quantitation of these important biopol ymers. Columns prepared by the procedure described in this study were recently applied to investigate proteolytic digests using different enzymes [39]. An investigation of the structural and biochemical aspects of femtin using these columns will be published elsewhere [40].…”

Section: Resultsmentioning

confidence: 99%

Simple method for the preparation of highly efficient polymer‐coated capillary electrophoresis columns

Malik

Zhao

Lee

1993

J Microcolumn Separations

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Abstract.A simple method for the preparation of polymer-coated fused silica capillary columns for electrophoresis is presented. It is shown that the static coating technique used in GC and SFC can be conveniently utilized for the preparation of highly efficient capillary electrophoresis columns. In this method, the coating solution contains appropriate proportions of three ingredients: a polymer, a surface derivatization reagent, and a crosslinking reagent dissolved in a suitable low-boiling solvent. After coating, the column is subjected to heat treatment to immobilize the polymer film. Simplicity, rapidity, high column efficiency, batch-to-batch and run-to-run reproducibility, and long column life-time are advantageous features of the new method. Column efficiencies of as high as 1.2 million theoretical plates were achieved for cytochrome c on a 96 cm long Ucon 75 H-90,000 coated column. Efficient electrophoretic separation of five cytochrome c proteins was achieved on a Superox 4 coated column. The possibility of CE separation of histones on the new columns was also explored.

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“…To determine protein MWs, He and Yeung [51] used PEO as a sieving matrix in an eight-capillary array electrophoresis system. Zhao et al [52,53] obtained highquality electrophoretic separations of basic proteins and peptides using fused-silica capillaries covalently coated with PEO/PEG.…”

Section: Peo/peg-coated Surfacesmentioning

confidence: 99%

Permanent surface modification of polymeric capillary electrophoresis microchips for protein and peptide analysis

Liu¹,

Lee²

2006

Electrophoresis

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102

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Because of their surface heterogeneity, proteins readily adsorb on polymeric substrates via various interactions, which adversely affects the performance of polymeric microfluidic devices in electrophoresis-based protein/peptide analysis. Therefore, it is necessary to use surface modification techniques such as dynamic coating or more complicated permanent surface modification, which has broader application and better performance, to render the polymeric microchannels protein-resistant. This manuscript is a review of the surface chemistry of microfluidic devices used for electrophoretic separations of proteins and peptides. The structural complexity of proteins as it relates to adsorption is described, followed by a review of the mechanisms and structural characteristics of protein-resistant surfaces. Permanent surface modification techniques used in grafting protein-resistant materials onto the surfaces of electrophoresis microchannels fabricated from polymer substrates are summarized and successful examples are presented.

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Separations of proteins and proteolytic digests of proteins by capillary electrophoresis on superox‐coated open tubular columns

Cited by 13 publications

References 31 publications

Surface-Modified Poly(methyl methacrylate) Capillary Electrophoresis Microchips for Protein and Peptide Analysis

Surface-Modified Poly(methyl methacrylate) Capillary Electrophoresis Microchips for Protein and Peptide Analysis

Simple method for the preparation of highly efficient polymer‐coated capillary electrophoresis columns

Permanent surface modification of polymeric capillary electrophoresis microchips for protein and peptide analysis

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