In situ photo-immobilised pH gradient isoelectric focusing and zone electrophoresis integrated two-dimensional microfluidic chip electrophoresis for protein separation

2015 ◽  
Vol 182 (13-14) ◽  
pp. 2321-2328 ◽  
Author(s):  
Fengmin Lin ◽  
Shiyong Yu ◽  
Le Gu ◽  
Xuetao Zhu ◽  
Jianshe Wang ◽  
...  
Keyword(s):  
Isoelectric Focusing ◽  
Microfluidic Chip ◽  
Protein Separation ◽  
Ph Gradient ◽  
Two Dimensional ◽  
Zone Electrophoresis

Native Protein Separation by Isoelectric Focusing and Blue Gel Electrophoresis-Coupled Two-Dimensional Microfluidic Chip Electrophoresis

2014 ◽  
Vol 77 (19-20) ◽  
pp. 1339-1346 ◽  
Author(s):  
Ni Hou ◽  
Yu Chen ◽  
Shiyong Yu ◽  
Zongliang Quan ◽  
Chenhua Pan ◽  
...  
Keyword(s):  
Isoelectric Focusing ◽  
Microfluidic Chip ◽  
Gel Electrophoresis ◽  
Protein Separation ◽  
Two Dimensional ◽  
Native Protein

Focusing of Proteins in a Horseshoe Microchannel

2008 ◽  
Author(s):  
Jaesool Shim ◽  
Prashanta Dutta ◽  
Cornelius F. Ivory
Keyword(s):  
Capillary Electrophoresis ◽  
Electric Field ◽  
Isoelectric Focusing ◽  
Ph Gradient ◽  
Complex Geometries ◽  
Electrokinetic Phenomena ◽  
Double Peaks ◽  
Zone Electrophoresis ◽  
Carrier Ampholytes ◽  
Ph Range

Ampholyte based isoelectric focusing (IEF) simulation was conducted to study dispersion of proteins in a horseshoe microchannel. Four model proteins (pls = 6.49, 7.1, 7.93 and 8.6) are focused in a 1 cm long horseshoe channel under an electric field of 300 V/cm. The pH gradient is formed in the presence of 25 biprotic carrier ampholytes (ΔpK = 3.0) within a pH range of 6 to 9. The proteins are focused at 380 sec in a nominal electric field of 300 V/cm. Our numerical results show that the band dispersions of a protein are large during the marching stage, but the dispersions are significantly reduced when the double peaks start to merge. This rearrangement of spreading band is very unique compared to linear electrokinetic phenomena (capillary electrophoresis, zone electrophoresis or electroosmosis) and is independent of channel position and channel shape. Hence, one can perform IEF in complex geometries without incorporating hyperturns.

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

2005 ◽  
Author(s):  
Prashanta Dutta ◽  
Keisuke Horiuchi ◽  
Huanchun Cui ◽  
Cornelius F. Ivory
Keyword(s):  
Isoelectric Focusing ◽  
Microfluidic Chip ◽  
Soft Lithography ◽  
Fluorescent Protein ◽  
Ph Gradient ◽  
Resolving Power ◽  
Protein Species ◽  
Second Stage ◽  
On Chip ◽  
Bonding Technique

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.

Isoelectric focusing in serial immobilized pH gradient gels to improve protein separation in proteomic analysis

2005 ◽  
Vol 26 (16) ◽  
pp. 3185-3190 ◽  
Author(s):  
Slobodan Poznanovic ◽  
Gerhard Schwall ◽  
Helmut Zengerling ◽  
Michael A. Cahill
Keyword(s):  
Isoelectric Focusing ◽  
Proteomic Analysis ◽  
Protein Separation ◽  
Ph Gradient
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