Issue 11, 2013

Photopatterned free-standing polyacrylamide gels for microfluidic protein electrophoresis

Abstract

Designed for compatibility with slab-gel polyacrylamide gel electrophoresis (PAGE) reagents and instruments, we detail development of free-standing polyacrylamide gel (fsPAG) microstructures supporting electrophoretic performance rivalling that of microfluidic platforms. For the protein electrophoresis study described here, fsPAGE lanes are comprised of a sample reservoir and contiguous separation gel. No enclosed microfluidic channels are employed. The fsPAG devices (120 μm tall) are directly photopatterned atop of and covalently attached to planar polymer or glass surfaces. Leveraging the fast <1 h design–prototype–test cycle – significantly faster than mold based fabrication techniques – we optimize the fsPAG architecture to minimize injection dispersion for rapid (<1 min) and short (1 mm) protein separations. The facile fabrication and prototyping of the fsPAGE provides researchers a powerful tool for developing custom analytical assays. We highlight the utility of assay customization by fabricating a polyacrylamide gel with a spatial pore-size distribution and demonstrate the resulting enhancement in separation performance over a uniform gel. Further, we up-scale from a unit separation to an array of 96 concurrent fsPAGE assays in 10 min run time driven by one electrode pair. The fsPAG array layout matches that of a 96-well plate to facilitate integration of the planar free standing gel array with multi-channel pipettes while remaining compatible with conventional slab-gel PAGE reagents, such as staining for label-free protein detection. Notably, the entire fsPAGE workflow from fabrication, to operation, and readout uses readily available materials and instruments – making this technique highly accessible.

Graphical abstract: Photopatterned free-standing polyacrylamide gels for microfluidic protein electrophoresis

Supplementary files

Article information

Article type
Paper
Submitted
25 Feb 2013
Accepted
04 Apr 2013
First published
05 Apr 2013

Lab Chip, 2013,13, 2115-2123

Photopatterned free-standing polyacrylamide gels for microfluidic protein electrophoresis

T. A. Duncombe and A. E. Herr, Lab Chip, 2013, 13, 2115 DOI: 10.1039/C3LC50269D

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