Two-dimensional electrophoretic separation of proteins using poly(methyl methacrylate) microchips.

Full Abstract

The work presented herein describes highly efficient, two-dimensional (2D) electrophoretic separations of proteins in a PMMA-based microchip. Sodium dodecyl sulfate microcapillary gel electrophoresis (SDS micro-CGE) and micellar electrokinetic chromatography (MEKC) were used as the separation modes for the first and second dimension of the electrophoresis, respectively. The microchip was prepared by hot embossing into PMMA from a brass mold master fabricated via high-precision micromilling. The microchip incorporated a 30-mm SDS micro-CGE and a 10-mm MEKC dimension length. Electrokinetic injection and separation were used with field strengths of up to 400 V/cm. Alexa Fluor 633 conjugated proteins, ranging in size from 38 to 110 kDa, were detected using laser-induced fluorescence with excitation/emission at 633/652 nm. Average plate numbers (N) of 4.8 x 10(4) and 1.2 x 10(4) were obtained in the SDS micro-CGE and MEKC separation dimensions, respectively, for the investigated proteins corresponding to plate heights (H) of 0.62 and 0.87 microm. Effluents from the first dimension (SDS micro-CGE) were repetitively transferred into the second dimension every 0.5 s of run time in the first dimension with the electrophoresis run time in the MEKC dimension being 10 s. The 2D separation was performed on the investigated proteins in approximately 12 min and provided a peak capacity of approximately 1000.

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Author information

Author/s: Shadpour, Hamed (H); Soper, Steven A (SA);

Affiliation: Department of Chemistry and Center for BioModular Multi-Scale Systems, Louisiana State University, Baton Rouge, Louisiana 70803, USA.

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.

Journal: Analytical chemistry (Anal Chem), published in United States. (Language: eng)

Reference: 2006-Jun; vol 78 (issue 11) : pp 3519-27

Dates: Created 2006/06/01; Completed 2007/04/13;

PMID: 16737203, status: MEDLINE (last retrieval date: 12/26/2008)

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

Comments and Corrections

CommentIn: Anal Chem. 2006 Jun 1;78(11):3489. (PMID: 16802459)

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Electrophoresis, Gel, Two-Dimensional - methods
Polymethyl Methacrylate - chemistry

Protein Array Analysis - methods
Proteins - analysis

Associated Chemicals: Proteins (0) ; Polymethyl Methacrylate (9011-14-7)

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