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 .
ABSTRACT
5#. The hybridoma cell lines grew well after continuous culture for more than three months or after being stored in liquid nitrogen for six months, and the titers of secreting McAbs were stable.Conclusion:The McAbs against respiratory syncytial virus are obtained. It lay a foundation for early diagnosis and further study of RSV.
ABSTRACT
The two hybridoma cell lines secreting antikeratin monoclonal antibodies (McAb)were obtained from fusion between NS_1 myelema cells and spleen cells of BALB/cmice immunized with keratin prepared from human callus.The two cell lines(AF_2and AF_5)have a chromosome number of 97 and 96 respectively.By immunodiffusion,both AF_2 and AF_5 McAb belong to mouse IgG_1 subclass.They have been successivelypassaged for over six months in vitro,and can grow well in the peritoneal fluid ofBALB/c mice,both produce high titer of specific antibodies. To determine their specificity with keratin of epithelia,AF_2 and AF_2 McAbwere used to stain various human normal and neoplastic tissue sections by the ABC(Avidin-Biotin peroxidase complex)immunohistochemical techniques.The results showthat AF_2 and AF_5 McAb can stain a series of normal epithelial tissues(in epidermis,positive for cells above the basal layer)and epithelial tumor tissues.No staining wereobserved in nonepithelial normal and tumor tissues.These results suggest that the twoMcAb are quite specific for a definite number of cytokeratins and promise to be use-ful in the differential diagnosis of tumors.