Production of anti-human c-kit monoclonal antibodies by direct intra-spleen injection of DNA vaccine
- VernacularTitle:基因疫苗脾内免疫制备抗人c-kit单克隆抗体及其初步鉴定
- Author:
Lin SHI
;
Dashui HE
;
Chunling FENG
;
Xiangfei YUAN
;
Hao QU
;
Lihua HUANG
;
Liyan ZHANG
;
Dongmei WANG
;
Yi ZHANG
;
Yuguang ZHANG
- Publication Type:Journal Article
- Keywords:
C-kit;
Genetic immunization;
Monoclonal antibody
- From:
Chinese Journal of Immunology
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare anti-human c-kit monoclonal antibody(McAb) by genetic immunization in spleen,and to determine practicability of these means to produce McAbs based on the biological activity of anti-human c-kit antibody.Methods:Recombinant plasmid pcDNA3.1/c-kit extracellular domain was constructed by molecular cloning techniques,and was used to immunize BALB/c mice in spleen directly to prepare mAb against human c-kit by routine hybridoma technique.FASC、fluorescence microscope and Western blot were utilized to identify the prepared antibody.Results:c-kit extracellular region was cloned and insert pcDNA3.1 plasmid successfully.Three hybridoma cell lines 6C4、2C5 and 5D5 that secrete anti-human c-kit McAbs were obtained after using intra-spleen immunization with a DNA vaccine.The isotypes of these three antibodies were all IgM,and the epitopes were different with each other.Conclusion:The method of genetic immunization into spleen can be used to prepare anti-human c-kit monoclonal antibodies.
