Preparation and purification of monoclonal antibodies against human chorionic gonadotrophin
- VernacularTitle:人绒促性素单克隆抗体的制备与分离纯化
- Author:
Hong SHEN
;
Yu YI
;
Jianfeng MEI
;
Keyin ZHU
;
Min LI
;
Guoqing YING
- Publication Type:Journal Article
- Keywords:
human chorionic gonadotrophin;
monoclonal antibody;
hybridoma cell lines
- From:
Chinese Journal of Biochemical Pharmaceutics
2010;31(1):22-25
- CountryChina
- Language:Chinese
-
Abstract:
Purpose To prepare monoclonal antibody against hCG. Methods Balb/c mice were immunized with hCG, and spleen cells from the mice were fused with myeloma cells SP2/0 at ratio of 5:1. Positive clones were screened by indirect enzyme-linked-immunoadsorbent assay (ELISA) ,then the clones were subcloned by limiting dilution and amplified further. After intraperitoneal injection of hybridoma cells, mouse ascites antibody was prepared. Titres and specificity of the ascites antibody were identified and determined by indirect ELISA. The ascites were purified by protein A affinity chromatography. Results Two strains of hybridoma cell lines obtained could secrete MAb stably. The titre of MAb of cell culture supernate is more than 10~(-3), and the titre of prepared ascites MAb is more than 10~(-7).The purity of the obtained monoclonal antibody was more than 98% and recovery reached 75 % after purification. Conclusion The obtained two strains of hybridoma cell lines can secrete MAb stably. The monoclonal antibodies can be used in the research of early pregnancy and tumor diagnosis .
