Concentration and separation of proteins in microfluidic channels on the basis of transverse IEF

被引:98
作者
Macounová, K [1 ]
Cabrera, CR [1 ]
Yager, P [1 ]
机构
[1] Univ Washington, Dept Bioengn, Seattle, WA 98195 USA
关键词
D O I
10.1021/ac001013y
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The use of microfluidic channels formed by two electrodes made of gold or palladium to perform transverse isoelectric focusing (IEF) is presented as a means for continuous concentration and fractionation of proteins. The micro-channels were 40 mm long with an electrode gap of 1.27 mm and a depth of 0.354 mm. The properties of pH gradients formed as a result of the electrolysis of water were influenced by variation of parameters such as the initial pH, ionic strength, and flow rate. Transverse IEF in pressure-driven flow is demonstrated using bovine serum albumin in a single ampholyte buffer as well as in multiple-component buffers, Experimental results of protein focusing compare well to predictions of a mathematical model. Optimal conditions for efficient continuous fractionation of a protein mixture are summarized and discussed.
引用
收藏
页码:1627 / 1633
页数:7
相关论文
共 24 条
[1]   Formation of natural pH gradients in a microfluidic device under flow conditions: Model and experimental validation [J].
Cabrera, CR ;
Finlayson, B ;
Yager, P .
ANALYTICAL CHEMISTRY, 2001, 73 (03) :658-666
[2]  
Cabrera CR, 2001, ELECTROPHORESIS, V22, P355, DOI 10.1002/1522-2683(200101)22:2<355::AID-ELPS355>3.0.CO
[3]  
2-C
[4]   Evaluation of commonly used electrophoretic methods for the analysis of proteins and peptides and their application to biotechnology [J].
Chiou, SH ;
Wu, SH .
ANALYTICA CHIMICA ACTA, 1999, 383 (1-2) :47-60
[5]   SEPARATION OF 2 COMPONENTS OF HORSE MYOGLOBIN BY ISOELECTRIC-FOCUSING FIELD-FLOW FRACTIONATION [J].
CHMELIK, J ;
DEML, M ;
JANCA, J .
ANALYTICAL CHEMISTRY, 1989, 61 (08) :912-914
[6]   ISOELECTRIC-FOCUSING FIELD-FLOW FRACTIONATION - EXPERIMENTAL-STUDY OF THE GENERATION OF PH GRADIENT [J].
CHMELIK, J .
JOURNAL OF CHROMATOGRAPHY, 1991, 539 (01) :111-121
[7]   ISOELECTRIC-FOCUSING FIELD-FLOW FRACTIONATION .4. INVESTIGATIONS ON PROTEIN SEPARATIONS IN THE TRAPEZOIDAL CROSS-SECTION CHANNEL [J].
CHMELIK, J ;
THORMANN, W .
JOURNAL OF CHROMATOGRAPHY, 1992, 600 (02) :305-311
[8]   NONEQUILIBRIUM THEORY OF FIELD-FLOW FRACTIONATION [J].
GIDDINGS, JC .
JOURNAL OF CHEMICAL PHYSICS, 1968, 49 (01) :81-&
[9]   Adaptation of capillary isoelectric focusing to microchannels on a glass chip [J].
Hofmann, O ;
Che, DP ;
Cruickshank, KA ;
Müller, UR .
ANALYTICAL CHEMISTRY, 1999, 71 (03) :678-686
[10]  
HOLL MR, UNPUB ELECTROPHORESI