摘要
为制备人血管内皮生长因子(VEGF)单克隆抗体,制备并纯化了VEGF蛋白,通过免疫、细胞融合、筛选等方法获得了能够稳定分泌单克隆抗体的细胞株;采用小鼠腹腔接种法制备腹水,并用G蛋白(Protein G)亲和层析柱对抗体进行了纯化;采用间接ELISA及Western blot实验分别对纯化后的抗体进行了效价测定和特异性验证.结果表明:此方法可成功筛选出能稳定分泌VEGF单克隆抗体的杂交瘤细胞株,纯化后腹水抗体效价为1∶1 024;Western blot检验证实制备的单克隆抗体能特异性识别VEGF蛋白.
The aim of this research was to prepare VEGF monoclonal antibody. The recombinant VEGF protein was prepared and purified, and the hybridoma cell lines which secreted stable monoclonal antibodies against VEGF antigen was produced through immunization, cell fusion, screening and other methods. Antibody was isolated using Protein G column. The titer and specificity of monoclonal antibody was analyzed by indirect ELISA and Western blot respectively. Results showed that the hybridoma cell lines which secreted stable monoclonal antibodies against VEGF antigen were successfully screened, and the titer was 1 ： 1 024. Western blot analysis showed that the VEGF monoclonal antibody could recognize VEGF protein specifically.
出处
《东北师大学报（自然科学版）》
CAS
CSCD
北大核心
2013年第3期126-129,共4页
Journal of Northeast Normal University(Natural Science Edition)
基金
长春市科技发展计划项目(长科技合2010058号)
国家自然科学基金资助项目(81028010)
