Combinatorial screening of enzyme activity by using multiplexed capillary electrophoresis

被引:51
作者
Ma, LJ
Gong, XY
Yeung, ES [1 ]
机构
[1] US DOE, Ames Lab, Ames, IA 50011 USA
[2] Iowa State Univ, Dept Chem, Ames, IA 50011 USA
关键词
D O I
10.1021/ac000145o
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Efficient and comprehensive screening of enzyme activity was accomplished in a combinatorial away of 96 reaction microvials. Quantitation of the extent of the reaction at web-defined time intervals was achieved by using 96-capillary array electrophoresis coupled with a multiplexed absorption detector. Capillary electrophoresis provides high separation resolution to isolate the product from the reactants. Absorption detection provides universal applicability to combinatorial screening. For the conversion of NADH to NAD(+), the catalytic activity of LDH was confirmed to be the highest at pH 7. This scheme should be useful for high-throughput drug discovery, clinical diagnosis, substrate binding, as well as combinatorial synthesis.
引用
收藏
页码:3383 / 3387
页数:5
相关论文
共 17 条
[1]   ULTRAMICRO ENZYME ASSAYS IN A CAPILLARY ELECTROPHORETIC SYSTEM [J].
BAO, JM ;
REGNIER, FE .
JOURNAL OF CHROMATOGRAPHY, 1992, 608 (1-2) :217-224
[2]  
BORMAN S, 1999, CHEM ENG NEWS 0308, P33
[3]  
BOTTOMLEY RH, 1966, BLOOD, V29, P182
[4]   Accessing genetic information with high-density DNA arrays [J].
Chee, M ;
Yang, R ;
Hubbell, E ;
Berno, A ;
Huang, XC ;
Stern, D ;
Winkler, J ;
Lockhart, DJ ;
Morris, MS ;
Fodor, SPA .
SCIENCE, 1996, 274 (5287) :610-614
[5]   An absorption detection approach for multiplexed capillary electrophoresis using a linear photodiode array [J].
Gong, XY ;
Yeung, ES .
ANALYTICAL CHEMISTRY, 1999, 71 (21) :4989-4996
[6]  
KAPLAN A, 1988, CLIN CHEM
[7]   COMPENSATING FOR INSTRUMENTAL AND SAMPLING BIASES ACCOMPANYING ELECTROKINETIC INJECTION IN CAPILLARY ZONE ELECTROPHORESIS [J].
LEE, TT ;
YEUNG, ES .
ANALYTICAL CHEMISTRY, 1992, 64 (11) :1226-1231
[8]   DNA-FINGERPRINTING BY HYBRIDIZATION TO OLIGONUCLEOTIDE ARRAYS [J].
SOUTHERN, EM .
ELECTROPHORESIS, 1995, 16 (09) :1539-1542
[9]   Monitoring the reactions of single enzyme molecules and single metal ions [J].
Tan, WH ;
Yeung, ES .
ANALYTICAL CHEMISTRY, 1997, 69 (20) :4242-4248
[10]   RNA aptamers specifically interact with the prion protein PrP [J].
Weiss, S ;
Proske, D ;
Neumann, M ;
Groschup, MH ;
Kretzschmar, HA ;
Famulok, M ;
Winnacker, EL .
JOURNAL OF VIROLOGY, 1997, 71 (11) :8790-8797