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Dynamic Concentration and Separation by Isoelectric Focusing in Microfluidic Channel
양광석,( Philippe Clementz ),김도현 한국화학공학회 2007 화학공학의이론과응용 Vol.10 No.1
A major focus in biochemistry is to explore the remarkable range of functions of proteins, because they play an important role in nearly all biological process. Thus, nowadays, many efforts are engaged into new proteomic analysis tools. A trend in microfluidics is integration of these proteins exploration techniques on lab-on-chip for micro total analysis systems (μ-TAS). To handle very small volumes is the promise of a great breakthrough in medical analysis. To separate and concentrate proteins in complex mixture, the Process Analysis Laboratory is developing an easy-tofabricatesolution integrated in a microfluidic device. In this study, we are targeting proteins concentration and separation of proteins on the basis of isoelectric focusing (IEF). We have designed and developed a polydimethylsiloxane (PDMS) microchip where dynamic pH gradient is successfully generated by varying the pH of buffer solution and biological particles (RFP-SBD Fusion proteins) in buffer solution undergo concentration and separation in a channel formed by electrodes.
Free-flow isoelectric focusing microfluidic device with glass coating by sol–gel methods
Kwang Suk Yang,Philippe Clementz,Tae Jung Park,Seok Jae Lee,Jong Pil Park,김도현 한국물리학회 2009 Current Applied Physics Vol.9 No.2
A device for microfluidic free-flow isoelectric focusing (μ-FFIEF) was fabricated in a glass-coated PDMS structure. The microfluidic channel was made by a PDMS replica molding and standard soft photolithography technique. For the prefractionation and preconcentration of a protein mixture, we patterned palladium electrodes on the glass substrate and coated with TEOS–sol to enhance the bonding with PDMS. The PDMS microchannel was coated with glass by flushing TEOS–sol and annealing at 100 ℃. The optimum focusing condition was obtained by pH indicator with ampholyte solution made with MES. Model proteins, RFP and EGFP, were successfully concentrated around 214 ㎛ and 357 ㎛ from the anode, respectively. The optimum focusing condition in the μ-FFIEF device was an electric potential of 1.5 V/cm and pH 6.0. The focusing time in the channel without glass coating was approximately 30 s, but the breakdown of the electrode by electrolysis started immediately when the electric field was applied. However, the glass-coated device was kept intact under continuous application of voltage giving a stable pH gradient, but with twice the focusing time.