摘要
本研究采用亲和毛细管电泳法(Affinity capillary electrop Horesis,ACE)对维拉帕米与牛血清白蛋白(Bovine Serum Albumin,BSA)的相互作用进行研究。以BSA为样品,以添加不同浓度药物的磷酸盐缓冲液(p H7.4,20 mmol·L^(-1))为背景缓冲溶液,采用以药物为添加剂的ACE方法,分别在25~oC和37~oC下进行毛细管电泳分离。根据各组分迁移时间,由Scatchard方程计算两个温度下药物与蛋白的结合常数K_b。实验结果表明,在ACE法中,随着药物浓度的增大,BSA与内标物的电泳迁移时间差值变大,由Scatchard方程计算得到维拉帕米与BSA在25~oC和37~oC时的K_b值分别为4.28×10~3M^(-1),ΔG为-20.73 k J·mol^(-1),ΔH为26.03 k J·mol^(-1),ΔS为157 J·mol^(-1)。在37℃时K_b值为6.43×10~3M^(-1),ΔG为-22.82 k J·mol^(-1),ΔH为26.03 k J·mol^(-1),ΔS为157 J·mol^(-1)。
Affinity capillary electrophoresis(ACE)was used to study the interaction between verapamil and bovine serum albumin(BSA).BSA was used as the sample.Phosphate buffer(pH7.4,20 mmol·L-1)was added as background buffer with different concentrations of drug.The ACE method with drug as additive was used for capillary electrophoresis separation at 25 o C and 37 o C,respectively.According to the migration time of each component,the binding constant K b of the drug and the protein at two temperatures was calculated from the Scatchard equation.The experimental results showed that in the ACE method,as the drug concentration increased,the difference between the electrophoretic migration time of the BSA and the internal standard became larger.The Scatchard equation was used to calculate the K b of Verapamil and BSA at 25 o C and 37 o C.The values were 4.28×10 3 M-1,ΔG was-20.73 kJ·mol-1,ΔH was 26.03 kJ·mol-1,andΔS was 157 J·mol-1.The K b value at 37℃was 6.43×10 3M-1,ΔG was 22.82 kJ·mol-1,ΔH was 26.03 kJ·mol-1,andΔS was 157J·mol-1.
作者
张剑
张博
唐一梅
匡元元
刘春叶
ZHANG Jian;ZHANG Bo;TANG Yi-mei;KUANG Yuan-yuan;LIU Chun-ye(Xi'an Medical University,School of Pharmacy,Xi’an 710021,China)
出处
《化学研究与应用》
CAS
CSCD
北大核心
2018年第10期1697-1702,共6页
Chemical Research and Application
基金
国家自然科学基金项目(81202492)资助
国家级大学生创新创业训练计划项目(2017DC-10
201711840015)资助
陕西省大创项目(2317)资助
校级大创项目(2017DC-10
2017DC-68)资助
西安医学院2017年国家基金培育项目(2017GJFY06)资助
关键词
维拉帕米
牛血清白蛋白
亲和毛细管电泳法
结合常数
verapamil
bovine serum albumin
affinity capillary electrophoresis
binding constant
