title: Protein separation using free-flow electrophoresis microchip etched in a single step.
abstract: A one-step etching method was developed to fabricate glass free-flow electrophoresis microchips with a rectangle separation microchamber (42 mm-long, 23 mm-wide and 28 microm-deep), in which two glass bridges (0.5 mm-wide) were made simultaneously to prevent bubbles formed by electrolysis near the Pt electrode from entering the separation chamber. By microchip free-flow zone electrophoresis, with 200 V voltage applied, the baseline separation of three FITC labeled proteins, ribonuclease B, myoglobin and beta-lactoglobulin, was achieved, with resolution over 1.78. Furthermore, with 2.5 mM Na(2)SO(4) added into the electrode buffer to form higher electrical field strength across separation microchamber than electrode compartments, similar resolution of samples was achieved with the applied voltage decreased to 75 V, which could obviously decrease Joule heat during continuous separation. All these results demonstrate that the free-flow electrophoresis microchip fabricated by one-step etching method is suitable for the continuous separation of proteins, which might become an effective pre-fractionation method for proteome study.
authors.affiliation: Key Lab of Separation Science for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
authors.forename: Pingli
authors.initials: P
authors.lastname: Wang
authors.forename: Lihua
authors.initials: L
authors.lastname: Zhang
authors.forename: Yichu
authors.initials: Y
authors.lastname: Shan
authors.forename: Yongzheng
authors.initials: Y
authors.lastname: Cong
authors.forename: Yu
authors.initials: Y
authors.lastname: Liang
authors.forename: Bin
authors.initials: B
authors.lastname: Han
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liang
authors.forename: Yukui
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D046209:Electrophoresis, Microchip / Q000379:methods*; D007782:Lactoglobulins / Q000302:isolation & purification*; D009211:Myoglobin / Q000302:isolation & purification*; D012260:Ribonucleases / Q000302:isolation & purification*
chemical_list: D007782:Lactoglobulins; D009211:Myoglobin; D012260:Ribonucleases; C029600:ribonuclease B