用多抗原肽(MAP)免疫制备单克隆抗体的研究  

Making Monoclonal Antibodies Specific for Native Proteins by Synthetic Multiple Antigenic Peptide

作　　者：丛进阳[1] 盖晓东[1] 

机构地区：[1]清华大学

出　　处：《生物工程学报》1997年第3期313-316,共4页Chinese Journal of Biotechnology

基　　金：国家自然科学基金

摘　　要：用多抗原肽（ＭＡＰ）免疫制备单克隆抗体的研究丛进阳盖晓东（清华大学北京１０００８４）目前制备单克隆抗体杂交瘤细胞株的传统方法是用纯度很高的天然蛋白作为抗原来免疫动物。此方法虽然成功率很高，但提取和纯化作为抗原用的高纯蛋白确非常困难，在某些情况下甚至是...A method for making hybridoma cell lines producing monoclonal antibodies specific for native protein by immunization of mice with synthetic multiple antigenic peptides (MAP) has been introduced. Fourteen amino acid sequence AAQYNPEPPPPRTH was selected from the small subunits of calpains according to its hydrophilicity index, and linked to lysine core with 8 branching sites to form a MAP. The MAP was used as an antigen for immunization of BALB/c mice. The mice strongly responded in two months after 3 injections. The antisera taken from the mice showed specific reaction with 28 kDa small subunits of calpains in Western blotting assay.Two fusions were made. As a result, four cell lines belonging to IgG subclasses were obtained. The investigation of these 4 cell lines by ELISA and Western blotting indicated that two cell lines (2C1 and 7B2) had specific reactions with 28 kDa subunits of both μ calpain and m calpain. Making monoclonal antibodies against native proteins by immunization of mice with synthetic MAP has showed usefulness of this method. The principles for selection of peptides suitable for making monoclonal antibodies against the native protein from which the peptide was selected, and the advantages of MAP were discussed briefly.

关 键 词：单克隆抗体 多抗原肽 制备 免疫学 

分 类 号：R392－33[医药卫生—免疫学]