摘要
目的制备半胱氨酸和甘氨酸富集蛋白2(Cysteine and glycine-rich protein 2,CRP2)的单克隆抗体,探讨CRP2蛋白的生物学功能。方法利用重组CRP2蛋白为免疫原,免疫BALB/C小鼠,取免疫小鼠的脾细胞和同系小鼠的骨髓瘤细胞SP2/0进行常规融合,通过间接ELISA筛选和有限稀释克隆化,获得鼠抗CRP2蛋白单克隆抗体的杂交瘤细胞株,通过ELISA,Western-blot和免疫共沉淀实验等方法对其特性进行鉴定。结果成功地建立了2株稳定分泌抗CRP2蛋白的单克隆抗体杂交瘤细胞株,分别命名为F001和F002。两株单克隆抗体与重组抗原有较强的特异性反应。同时,两株单克隆抗体通过Western-Blot实验都能特异性地结合真核细胞内源性的CRP2蛋白,并且,两株抗体都可以应用于免疫共沉淀实验。结论获得了效价高、特异性好的CRP2蛋白的单克隆抗体,为CRP2的生物学功能研究奠定了基础。
Objective To obtain monoclonal antibodies against cysteine and glycine- rich protein 2 （CRP2） for further study of the biological function of CRP2 protein. Methods Balb/c mice were immunized with recombinant CRP2. Hybridoma cell lines secreting monoclonal antibodies against CRP2 were screened by regular cell fusion and subcloning approach. The specificities of these monoclonal antibodies were determined by ELISA, Western blotting and immunoprecipitation. Results Two hybridoma cell lines （F001 and F002） stable in secreting specific monoclonal antibodies were successfully obtained. They specifically bind to recombinant CRP2 and endogenous CRP2 proteins proved by ELISA, Western blotting. In addition, these two monoclonal antibodies could be precipitated the endogenous CRP2 protein by immunopre- cipitation assay. Conclusion Monoclonal antibodies against CRP2 with high titers and specificity have been successfully prepared, providing foundation for further study of CRP2 protein.
出处
《广东医学》
CAS
CSCD
北大核心
2008年第2期201-203,共3页
Guangdong Medical Journal
基金
国家高技术研究发展计划(863计划)项目(编号:2006AA02A311)
