title: Microfluidic two-dimensional separation of proteins combining temperature gradient focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
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.
authors: Department of Mechanical and Mechatronics Engineering, University of Waterloo, 200 University Ave. West, Waterloo, Ontario Canada, N2L 3G1.
authors: Seyed Mostafa
authors: SM
authors: Shameli
authors: Department of Mechanical and Mechatronics Engineering, University of Waterloo, 200 University Ave. West, Waterloo, Ontario Canada, N2L 3G1.
authors: Carolyn L
authors: CL
authors: Ren
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation*; D004867:Equipment Design; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D013696:Temperature
chemical_list: D011506:Proteins