Dynamic coating using methylcellulose and polysorbate 20 for nondenaturing electrophoresis of proteins on plastic microchips

A dynamic coating using methylcellulose (MC) and a nonionic detergent (polysorbate 20) was developed, which controlled protein adsorption onto the surface of microchannels on a microchip made of poly(methyl methacrylate) (PMMA). Optimum concentration of polysorbate 20 in combination with the range o...

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Bibliographic Details
Published in:Electrophoresis 2007-03, Vol.28 (5), p.830-836
Main Authors: Mohamadi, Mohamad Reza, Mahmoudian, Laili, Kaji, Noritada, Tokeshi, Manabu, Baba, Yoshinobu
Format: Article
Language:English
Subjects:
Adsorption
Animals
Buffers
Cattle
Dynamic coating
Electrophoresis, Microchip - methods
Glucan 1,4-alpha-Glucosidase - analysis
Methylcellulose - chemistry
PMMA microchip
Polymethyl Methacrylate - chemistry
Polysorbates - chemistry
Protein electrophoresis
Serum Albumin, Bovine - analysis
Surface Properties
Trypsin Inhibitors - analysis
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Summary:A dynamic coating using methylcellulose (MC) and a nonionic detergent (polysorbate 20) was developed, which controlled protein adsorption onto the surface of microchannels on a microchip made of poly(methyl methacrylate) (PMMA). Optimum concentration of polysorbate 20 in combination with the range of MC concentrations controlled the protein adsorption onto the microchannel surface, and increased the solubility of the protein samples while facilitating the injection of high concentrations of MC solutions into the microchannels. Higher concentrations of nonionic detergent increased the EOF mobility as opposed to the electrophoretic mobility and caused the electrophoresis to fail. Nondenaturing microchip electrophoresis of protein samples with molecular masses ranging from 20 to 100 kDa were completed in 100 s. Also, successful separation of a BSA sample and its complex with anti‐BSA mAb (˜220 kDa) was achieved on a PMMA microchip. The separation exhibited high reproducibility in both migration time (RSD = 1%) and peak area (RSD = 10–15%).
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.200600373