摘要
根据生物素标记反应的不可逆性,设计了一种基于磁性分离的快速纯化方法.该方法使用固定在磁性粒子上的牛血清白蛋白(BSA)与生物素标记甲状腺素(T4)反应后过量的活性生物素试剂继续反应,形成磁性粒子-BSA-生物素复合物,再通过磁性分离可快速除去复合物.实验中用酶免疫分析方法检测过量的活性生物素试剂和磁性粒子的反应程度,并用竞争抑制反应证明了经磁性分离得到的上清液为高纯度生物素标记的T4.该纯化方法解决了小分子物质生物素标记后难以用传统方法纯化的问题,操作快速、简单,特别适用于小分子生物物质进行生物素标记后的纯化.
According to the irreversibility of biotin labeling, a quick method was designed for the purification of biotin labeling small molecule by magnetic separation. Bovine serum albumin （BSA） coated on the magnetic particles was used to react with the excessive active biotin reagent after T4 was biotinylated, forming magnetic particles -BSA-biotin complexes, which can be quickly removed by a magnet. Enzyme immunoassay was used to detect the reactive degree between excessive biotin reagent and magnetic particles. And the supernatant liquid obtained after magnetic separation was proved to be highly pure biotinylated T4 by a competitive inhibition reaction. The method solved the problem that small molecule after labeled by biotin is difficult to be purified, and can be operated quickly and simply. It is very efficient for the purification of biotinylated small molecule after labeling reaction.
出处
《上海交通大学学报》
EI
CAS
CSCD
北大核心
2005年第8期1379-1382,共4页
Journal of Shanghai Jiaotong University
基金
国家高技术研究发展计划(863)项目(2002AA302210)
关键词
生物素
磁性分离
牛血清白蛋白
磁性粒子
酶联免疫分析
竞争抑制反应
biotin
magnetic separation
bovine serum albumin（BSA）
magnetic particles
enzyme immunoassay
competitive inhibition reaction
