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STUDIES ON COUNTERACTING CHROMATOGRAPHIC electrophoresis AND ITS UTILIZATION FOR PROTEIN PURIFICATION

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Chinese Journal of Chemical Engineering 1996年第1期4卷 14-21页

作者：高瑞昶金曼蓉王世昌天津大学化学工程研究中心天津天津第二医学院药学系天津

A counteracting chromatographic electrophoresis(CACE)separation device has been success-fully built and tested.Experiments are performed with protein mixtures of BSA,Hg,Mg and Cyto-cto study the separation performances,concentration profiles,start-up methodology as well as influ-ences of pH and ionic strength on the separation process.Mathematical models based on the fluiddynamics of the porous medium are proposed.Experiments indicate that the CACE process has agood commercial potential in protein engineering and fine chemicals purification.

关键词： chromatographic electrophoresis proteins preparative separation porous medium

CONTINUOUS SEPARATION OF PROTEINS BY HYDROPHILIC POLYMER SHIELDED MULTICHANNEL FLOW electrophoresis

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Chinese Journal of Chemical Engineering 1997年第2期5卷 49-54页

作者：刘铮赵寅沈忠耀丁富新袁乃驹 Department of Chemical Engineering Tsinghua University

Continuous separation of protein mixtures by multichannel flow electrophoresis(MFE)wascarried out in a 5-compartment electrolyzer partitioned by membranes.Polyvinyl alcohol(PVA),polyethylene glycol 4000(PEG 4000)and polyvinylpyrrolidene K30(PVP K30)were applied to theMFE as shielding polymers to prevent protein adsorption on the polyethersulfone microfiltrationmembrane,which was used to space the central compartment and the elution compartments,duringthe electrophoresis process.The effects of polymer concentration on protein transmembrane flux wereexamined.It was found that PVA,PEG 4000 and PVP K30 greatly reduced protein adsorpton on themembrane surface.Continuous separations of bovine serum albumin(BSA)and hemoglobin(HBB)mix-ture in the presence of PEG 4000 yielded 26.6mg BSA 40.4mg HBB per hour.These results haveshown a high potential of scaling up MFE for large scale separation and purification ofbiomolecules.

关键词： electrophoresis multichannel flow electrophoresis

Direct fabrication of a microfluidic chip for electrophoresis analysis by water-assisted femtosecond laser writing in porous glass

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Chinese Optics Letters 2013年第7期11卷 57-60页

作者：居永凤刘昌宁廖洋柳洋张龙沈应龙陈丹平程亚 Shanghai Institute of Optics and Fine Mechanics Chinese Academy of Sciences University of Chinese Academy of Sciences

We report the direct fabrication of a microfluidic chip composed of two high-aspect ratio microfluidic channels with lengths of 3.5 cm and 8 mm in a glass substrate by femtosecond laser micromachining. The fabrication mainly consists of two steps： 1） writing microchannels and microchambers in a porous glass by scanning a tightly focused laser beam; 2） high-temperature annealing of the glass sample to collapse all the nanopores in the glass. Migration of derivatized amino acids is observed in the microfluidic channel by applying electric voltage across the long-migration microchannel.

关键词： Amino acids Aspect ratio electrophoresis Fabrication Fluidic devices Microchannels Substrates

Application of liquid pre-column capillary electrophoresis technique to the study of interaction between drug enantiomers and human serum albumin

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Science China Chemistry 1999年第6期42卷 617-623页

作者：丁永生朱晓蜂林炳承 Dalian Maritime University Dalian China Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian China

Based on the chiral separation of several basie drugs, dimetindene, tetryzoline, theodrenaline and verapamil, the liquid pre-colunm capillary electrophoresis (LPC-CE) technique was established. It was used to determine free concentrations of drug enantiomers in mixed solutions with human serum albumin (HSA). To prevent HSA entering the CE chiral separation zone, the mobility differences between HSA and drugs under a specific pH condition were employed in the LPC. Thus, the detection confusion caused by protein was totally avoided. Further study of binding constants determination and protein binding competitions was carried out. The study proves that the LPC technique could be used for complex media, particularly the matrix of protein coexisting with a variety of drugs.

关键词： capillary electrophoresis liquid pre-column binding constant chiral drug human serum albumin.

Capillary electrophoresis of FITC labeled amino acids with laser-induced fluorescence detection

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Science China Chemistry 1999年第6期42卷 663-669页

作者：党福全陈义 Center for Molecular Science Institute of Chemistry Chinese Academy of Sciences Beijing China

FITC labeled amino acids have been separated using a home-huilt capillary electrophoresis with a laserinduced fluorescence detection (CE-LIF) system. Seventeen peaks can now be generated from the twenty common amino a... 详细信息

关键词： capillary electrophoresis with laser-induced fluorescence amino acids fuorescein isothiocyanate (FITC).

Establishment and application of a new method for the determination of kinetic parameters by plug-plug kinetic capillary electrophoresis (ppKCE)

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Science China Chemistry 2008年第11期51卷 1087-1092页

作者： XIONG CaiQiao XIA ZhiNing HUANG Rui CHEN Hua XU Pan College of Chemistry and Chemical Engineering Chongqing UniversityChongqing 400044China College of Biological Engineering Chongqing UniversityChongqing 400044China

A novel plug-plug kinetic capillary electrophoresis method was established, which can be used to si-multaneously determine the kinetic parameters kon and koff in interaction systems. The method is comparatively simple and some restrictions in conventional ppKCE methods can be effectively avoided. The requirements for resolution and detection sensitivity in this method are much lower than those of conventional ppKCE. The successful determination of the kinetic parameters and the binding constant Kb between citalopram and BSA showed availability of this method. The results were confirmed by us-ing the time ratio method. The application field of kinetic capillary electrophoresis can be expanded with this new method.

关键词： interaction capillary electrophoresis plug-plug kinetic capillary electrophoresis (ppKCE) kinetic parameter

Separation of acidic and basic proteins by capillary electrophoresis using gemini surfactants and gemini-capped nanoparticles as buffer additives

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Science China Chemistry 2009年第10期52卷 1666-1676页

作者： LIU Qian LI YanQing YANG YanMin YAO ShouZhuo State Key Lab of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical EngineeringHunan UniversityChangsha 410082China

This paper demonstrated simultaneous separation of acidic and basic proteins using cationic gemini surfactants as buffer additives in capillary electrophoresis. We showed that even at a low concentration (0.1 mmol·L-1) of alkanediyl-α,ω-bis(dimethyloctadecylammonium bromide) (18-s-18), the wall adsorption of both acidic and basic proteins could be effectively suppressed under acidic conditions. Smaller micelle size (e.g., s=5-8) is more effective for the separation of acidic proteins than larger micelle size (e.g., s10). Varying the spacer length of gemini surfactants can influence the electrophoretic mobility and selectivity of proteins to achieve the desired separation. Under the optimized conditions, RSDs of the migration time were less than 0.8% and 2.2% for run-to-run and day-to-day assays, respectively, and protein recoveries ranged from 79% to 100.4%. Furthermore, we also investigated the use of gemini surfactant-capped gold nanoparticles (gemini@AuNPs) as buffer additives in protein separation. Introduction of AuNPs into the buffer shortened the analysis time and slightly improved the separation efficiencies. Finally, we presented the applications of this method in the analysis of bio-logical samples, including plasma, red blood cells and egg white.

关键词： gemini surfactant protein separation capillary electrophoresis gold nanoparticle

Construction and evaluation of a novel end-column amperometric detection system for electrophoresis microchips

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Science China Chemistry 2010年第1期53卷 250-256页

作者： CHEN ZhiFeng1,GAO YunHua2,WANG Li3,CHU XiaoGang1 1Chinese Academy of Inspection and Quarantine,Beijing 100123,China 2Technical Institute of Physics and Chemistry,Chinese Academy of Sciences,Beijing 100190,China 3Beijing Institute of Labour Protection,Beijing 100054,China Chinese Academy of Inspection and Quarantine Beijing China Technical Institute of Physics and Chemistry Chinese Academy of Sciences Beijing China Beijing Institute of Labour Protection Beijing China

A set of integrated end-column amperometric detection system has been developed,onto which an electrophoresis microchip can be conveniently integrated.Finely machined by a piece of transparent organic glass,the system consists of an electrophoresis microchip platform and an amperometric detection reservoir,in which the microchip can be fixed onto the platform by microchip grooves and with stainless steel fixture.Each detection electrode can be directly fixed in the amperometric detection reservoir by screws and nuts.With dopamine as the model analyte,we take platinum disc electrodes with different diameters of 100 μm,300 μm and 500 μm and a carbon fiber electrode with the diameter of 240 μm as the working electrode,all of which accomplish sensitive detection.The detection parameters of the system are optimized with the carbon fiber electrode.The detection results show that in the electrochemical cleaning procedure,the relative deviations of 3.2% and 0.5% for the peak current and retention time,respectively,can be obtained for the successive detections of 100 μM dopamine,and the limit of detection for dopamine can reach 0.4 μM(S/N = 3).This system is small,but its performance of detection is stable and sensitive,and the replacement of its working electrodes is convenient,so it is very suitable as a universal platform of end-column amperometric detection for electrophoresis microchips.

关键词： electrophoresis microchip amperometric detection dopamine evaluation

NEW APPROACH TO MINIMIZE DISPERSION INDUCED BY TURN IN CAPILLARY electrophoresis CHANNEL FLOWS

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Applied Mathematics and Mechanics(English Edition) 2005年第6期26卷 685-690页

作者：李志华林建忠聂德明 Department of Mechanics State Key Laboratory of Fluid Power Transmission and Control Zhejiang University Zhejiang University of Science and Technology Hangzhou 310012P.R.China

The mechanism of dispersion induced by turn in the capillary electrophoresis channel flows was analyzed firstly. Then the mathematical model of electroosmotic flow is built, and the dispersion of the flow, with different distribution of charge at inner and outer wall in the turns, was simulated numerically using the finite differential method. A new approach of altering the distribution of charge at inner and outer wall in the turns was presented, based on the computational results, to minimize the dispersion induced by turn. Meanwhile, an optimization algorithm to analyze the numerical results and determine the optimal distribution of charge in the turns was also developed. It is found that the dispersion induced by turn in the capillary electrophoresis channel flows could be significantly suppressed by this approach.

关键词： micro-flow electrophoresis electroosmotic dispersion turn