Microfluidic two-dimensional separation of proteins combining temperature gradient focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Analytical Chemistry
Seyed Mostafa Shameli, Carolyn L Ren

Abstract

A two-dimensional separation system is presented combining scanning temperature gradient focusing (TGF) and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) in a PDMS/glass microfluidic chip. Denatured proteins are first focused and separated in a 15 mm long channel via TGF with a temperature range of 16-47 °C and a pressure scanning rate of -0.5 Pa/s and then further separated via SDS-PAGE in a 25 mm long channel. A side channel is designed at the intersection between the two dimensions to continuously inject SDS into the gel, allowing SDS molecules to be compiled within the focused bands. Separation experiments are performed using several fluorescently labeled proteins with single point detection. Experimental results show a dramatic improvement in peak capacity over one-dimensional separation techniques.

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Citations

Jul 20, 2017·Electrophoresis·Zhichao WangAdrienne R Minerick
Aug 13, 2017·Electrophoresis·Yuanyuan LiuChao Yan
Dec 8, 2017·The Analyst·I Rodríguez-RuizJ A Gavira
Sep 10, 2019·ACS Applied Materials & Interfaces·Xuemin WangCraig A Aspinwall
Aug 22, 2017·Analytical Chemistry·Philippe Abdel-SayedAmy E Herr

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