The use of biotinylated monoclonal antibodies and streptavidin affinity chromatography to isolate herpesvirus hydrophobic proteins or glycoproteins.
The use of biotinylated monoclonal antibodies and streptavidin affinity chromatography to isolate herpesvirus hydrophobic proteins or glycoproteins.
复制标题
使用生物素化单克隆抗体和链霉亲和素亲和层析分离疱疹病毒疏水蛋白或糖蛋白。
DOI：
10.1016/0003-2697(87)90123-0
复制
发表时间：
1987
影响因子：
2.9
通讯作者：
Stinski,MF
中科院分区：
文献类型：
--
作者：
Gretch,DR;Suter,M;Stinski,MF
以链霉亲和素/生物素为基础的免疫亲和系统被优化为从粗制的感染细胞裂解物中分离疱疹病毒(人巨细胞病毒)的即刻早期蛋白或晚期糖蛋白。ϵ氨基生物素替代一抗的生物素化效果优于糖基生物素替代。一抗的生物素化效果优于二抗。用约8M生物素/米抗体的生物素替代可最大限度地回收病毒抗原。基于链霉亲和素/生物素的免疫亲和系统可以制备相对纯净的病毒抗原，可用于功能、免疫学或结构研究。
A streptavidin/biotin-based immunoaffinity system was optimized to isolate herpesvirus (human cytomegalovirus) immediate early proteins or late glycoproteins from crude infected cell lysates. Biotinylation of the primary antibody by biotin substitution of the ϵ amino groups was superior to biotin substitution of sugar residues. Biotinylation of the primary antibody was superior to that of a secondary antibody. A biotin substitution of approximately 8 m biotin/m antibody allowed for maximal recovery of viral antigens. The streptavidin/biotin-based immunoaffinity system can allow for relatively pure preparations of viral antigens that may be used for functional, immunological, or structural studies.