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Multistage Isoelectric Focusing: A Novel On-Chip Bio-Separation Technique

[+] Author Affiliations
Prashanta Dutta, Keisuke Horiuchi, Huanchun Cui, Cornelius F. Ivory

Washington State University

Paper No. IMECE2005-79978, pp. 229-234; 6 pages
doi:10.1115/IMECE2005-79978
From:
  • ASME 2005 International Mechanical Engineering Congress and Exposition
  • Fluids Engineering
  • Orlando, Florida, USA, November 5 – 11, 2005
  • Conference Sponsors: Fluids Engineering Division
  • ISBN: 0-7918-4219-3 | eISBN: 0-7918-3769-6
  • Copyright © 2005 by ASME

abstract

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.

Copyright © 2005 by ASME

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