Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors

被引:71
作者
Heintz, J [1 ]
Hernandez, M [1 ]
Gomez, FA [1 ]
机构
[1] Calif State Univ Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90032 USA
关键词
affinity capillary electrophoresis; binding constants; partial-filling affinity capillary electrophoresis; vancomycin; carbonic anhydrase; enzymes; antibiotics;
D O I
10.1016/S0021-9673(99)00278-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This work evaluates the concept of a partial-filling technique in affinity capillary electrophoresis (ACE) using two model systems: vancomycin from Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). In this technique the capillary is first partially-filled with ligand followed by a sample of receptor and non-interacting standard and electrophoresed. Analysis of the change in the mobility ratio, M, of the receptor, relative to the non-interacting standard, as a function of the concentration of the ligand, yields a value for the binding constant. These values agree well with those estimated using other binding and ACE techniques. Data demonstrating the quantitative potential of this method is presented. (C) 1999 Elsevier Science BN. All rights reserved.
引用
收藏
页码:261 / 268
页数:8
相关论文
共 61 条
[1]   Post-capillary affinity detection of protein microheterogeneity in capillary zone electrophoresis [J].
Abler, JK ;
Reddy, KR ;
Lee, CS .
JOURNAL OF CHROMATOGRAPHY A, 1997, 759 (1-2) :139-147
[2]   Relationship between retention and effective selector concentration in affinity capillary electrophoresis and high-performance liquid chromatography [J].
Ahmed, A ;
Ibrahim, H ;
Pastore, F ;
Lloyd, DK .
ANALYTICAL CHEMISTRY, 1996, 68 (18) :3270-3273
[3]   Enantioseparation of local anaesthetic drugs by capillary zone electrophoresis with cyclodextrins as chiral selectors using a partial filling technique [J].
Amini, A ;
PaulsenSorman, U .
ELECTROPHORESIS, 1997, 18 (06) :1019-1025
[4]   Evaluation of association constants between drug enantiomers and human α1-acid glycoprotein by applying a partial-filling technique in affinity capillary electrophoresis [J].
Amini, A ;
Westerlund, D .
ANALYTICAL CHEMISTRY, 1998, 70 (07) :1425-1430
[5]   CATALYTIC ACTIVITY OF NATIVE ENZYMES DURING CAPILLARY ELECTROPHORESIS - AN ENZYMATIC MICROREACTOR [J].
AVILA, LZ ;
WHITESIDES, GM .
JOURNAL OF ORGANIC CHEMISTRY, 1993, 58 (20) :5508-5512
[6]   ULTRAMICRO ENZYME ASSAYS IN A CAPILLARY ELECTROPHORETIC SYSTEM [J].
BAO, JM ;
REGNIER, FE .
JOURNAL OF CHROMATOGRAPHY, 1992, 608 (1-2) :217-224
[7]   Guidelines in selecting ligand concentrations for the determination of binding constants by affinity capillary electrophoresis [J].
Bose, S ;
Yang, J ;
Hage, DS .
JOURNAL OF CHROMATOGRAPHY B, 1997, 697 (1-2) :77-88
[8]   MOLECULAR-BASIS FOR VANCOMYCIN RESISTANCE IN ENTEROCOCCUS-FAECIUM BM4147 - BIOSYNTHESIS OF A DEPSIPEPTIDE PEPTIDOGLYCAN PRECURSOR BY VANCOMYCIN RESISTANCE PROTEINS VANH AND VANA [J].
BUGG, TDH ;
WRIGHT, GD ;
DUTKAMALEN, S ;
ARTHUR, M ;
COURVALIN, P ;
WALSH, CT .
BIOCHEMISTRY, 1991, 30 (43) :10408-10415
[9]   Vacancy affinity capillary electrophoresis, a new method for measuring association constants [J].
Busch, MHA ;
Boelens, HFM ;
Kraak, JC ;
Poppe, H .
JOURNAL OF CHROMATOGRAPHY A, 1997, 775 (1-2) :313-326
[10]   Comparison of five methods for the study of drug-protein binding in affinity capillary electrophoresis [J].
Busch, MHA ;
Carels, LB ;
Boelens, HFM ;
Kraak, JC ;
Poppe, H .
JOURNAL OF CHROMATOGRAPHY A, 1997, 777 (02) :311-328