4.5 Article

Exceeding 20 000-fold concentration of protein by the on-line isotachophoresis concentration in poly(methyl methacrylate) microchip

Journal

ELECTROPHORESIS
Volume 30, Issue 18, Pages 3250-3256

Publisher

WILEY-BLACKWELL
DOI: 10.1002/elps.200900111

Keywords

BSA; Immunoassay; ITP concentration; PMMA microchip

Funding

  1. Ministry of Education, Culture, Sports, Science and Technology, Japan

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In this research, a simple on-line microchip gel electrophoresis with ITP was applied for the concentration and separation of BSA and its immunoassay complex with mAb in a single cross form PMMA microchip. We investigated the ITP concentration effect in PMMA MCE using combination of leading electrolytes, terminating electrolytes and other factors. We realized an ITP-based concentration and separation of BSA and its immunoassay complexes in standard cross-channel microchip gel electrophoresis, which exceeded 2000-fold concentration of BSA immunocomplex using Tris-H3PO4 as a leading electrolyte and Tris-gamma-amino butyric acid as a terminating electrolyte. in addition, we also realized concentration of BSA sample in water, which was more than 20 000-fold and was the result of the concentration effect from combining ITP and the sample stacking techniques.

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