Stepwise frontal affinity chromatography model for drug and protein interaction

被引：17

作者：

He, Xiaoshuang ^{[1
,2
]}

Sui, Yue ^{[1
,2
]}

Wang, Sicen ^{[1
,2
]}

机构：

[1] Xi An Jiao Tong Univ, Sch Pharm, 76 Yanta West Rd, Xian 710061, Shaanxi, Peoples R China

[2] Shaanxi Engn Res Ctr Cardiovasc Drugs Screening &, 76 Yanta West Rd, Xian 710061, Shaanxi, Peoples R China

来源：

ANALYTICAL AND BIOANALYTICAL CHEMISTRY | 2018年 / 410卷 / 23期

基金：

中国国家自然科学基金;

关键词：

Affinity chromatography; Stepwise frontal analysis model; Lineweaver-Burk plot; Equilibrium dissociation constants (K-d); CELL-MEMBRANE CHROMATOGRAPHY; HUMAN SERUM-ALBUMIN; HUMAN ALPHA(1)-ACID GLYCOPROTEIN; MASS-SPECTROMETRY; ALPHA(1A) ADRENORECEPTOR; BIOACTIVE COMPOUNDS; BINDING; RECEPTOR; IDENTIFICATION; MICROCOLUMNS;

D O I：

10.1007/s00216-018-1194-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Frontal affinity chromatography is an efficient technique that combines affinity interaction and high-performance liquid chromatography, and frontal analysis has been used in studying the interaction between drugs and proteins. Based on frontal analysis, stepwise frontal analysis has been established. The present study aimed to use the Lineweaver-Burk plot in stepwise frontal analysis by taking the weighted average of time data. Commercial human serum albumin (HSA) and alpha-1-acid glycoprotein (AGP) columns were used as an affinity column. Warfarin and digitoxin were chosen as model drugs for the HSA column, whereas verapamil and tamsulosin were selected as model drugs for the AGP column. The time data obtained by frontal analysis and stepwise frontal analysiswere compared, and the results revealed good correlation (r2 = 0.9946-0.9998). Frontal analysis and stepwise frontal analysis were also used to analyze the equilibrium dissociation constants (Kd) of model drugs on the HSA and AGP columns. The Kd values were compared with literature values, which revealed the same order of magnitude. These results illustrate that conversion of the time data is reasonable and feasible. The Lineweaver-Burk plot can be used in the stepwise frontal analysis model to study the characteristics of the interaction between drugs and proteins.

引用

页码：5807 / 5815

页数：9

共 36 条

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