苏州市莱顿科学仪器有限公司
2009/2/20 12:23:15毛细管电泳应用于测定牛血清白蛋白与脂质体的相互作用
厉红,屈锋,徐建栋,邓玉林
北京理工大学生命科学与技术学院, 北京 100081
收稿日期 2008-2-21 修回日期 2008-7-30 网络版发布日期 2008-7-30 接受日期 2008-7-30
摘要 建立了一种用毛细管电泳法检测牛血清白蛋白(BSA)与脂质体相互作用的分析方法。氧化指数的测定实验结果表明经过冷冻干燥的脂质体稳定性更好;毛细管电泳表征脂质体的电荷性质实验结果表明脂质体在pH 5.0~8.0的条件下呈电中性。在pH 7.0的条件下,以各种浓度的脂质体混悬液为电泳缓冲液,以0.8%二甲亚砜(DMSO)为内标,随着缓冲液中脂质体质量浓度从0增加到2.4 mg/mL,BSA的有效淌度从-2.232×10-4 cm2·V-1·s-1变化到-3.046×10-4 cm2·V-1·s-1;结合Scatchard分析,测得BSA与脂质体的结合常数为2.522×103(g/mL)-1。该方法简单、快速,为研究蛋白质与脂质体的相互作用提供了新的技术手段。
关键词 毛细管电泳 牛血清白蛋白 脂质体
Investigation on interaction between bovine serum albumin and liposome using capillary electrophoresis
LI Hong, QU Feng, XU Jiandong, DENG Yulin
School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, China
Abstract
A method for the investigation on the interaction between bovine serum albumin (BSA) and liposome using capillary electrophoresis was developed. The oxidation index showed that the liposomes after freeze-drying were more stable. The results obtained from the capillary electrophoretic analysis of liposome showed that liposome had no charge at pH 5.0-8.0. A series of liposome suspension at different concentrations with the internal marker of 0.8% dimethyl sulfoxide (DMSO) were introduced as the electrophoresis buffer at pH 7.0. Along with the liposome concentrations raised from 0 to 2.4 mg/mL, it was found that the effective mobility of BSA changed from -2.232×10-4 cm2·V-1·s-1 to -3.046×10-4 cm2·V-1·s-1. The binding constant between BSA and liposome was 2.522×103(g/mL)-1 calculated by Scatchard analysis. This method is simple and rapid, and provides a new technology for the investigation on the interactions between protein and liposome.
Key words capillary electrophoresis (CE) bovine serum albumin (BSA) liposome