This experimental study reports a method to increase the resolving power of isoelectric focusing (IEF) on a polymeric microfluidic chip. Microfluidic chip is formed on poly-di-methyl siloxane (PDMS) using soft lithography and multilayer bonding technique. In this novel bioseparation technique, IEF is staged by first focusing protein species in a straight channel using broad-range ampholytes and then refocusing segments of that first channel into secondary channels that branch out from the first one. Experiments demonstrated that three fluorescent protein species within a segment of pH gradient in the first stage were refocused in the second stage with much higher resolution in a shallower pH gradient. A serially performed two-stage IEF was completed in less than 25 minutes under particularly small electric field strength up to 100 V/cm.

Volume Subject Area:
Fluids Engineering
Topics:
Bonding, Electric fields, Lithography, Microfluidics, Plasma desorption mass spectrometry, Proteins, Resolution (Optics), Separation (Technology), Siloxanes
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