人绒毛膜促性腺激素单克隆抗体的研制

Generation and identification of monoclonal antibody against human chorionic gonadotrophin

目的获得针对人绒毛膜促性腺激素单克隆抗体。方法hCG蛋白免疫Balb/c小鼠,取其脾细胞与同系小鼠骨髓瘤细胞NS-1按8∶1比例融合,间接ELISA法筛选阳性克隆,有限稀释法进行克隆化培养;制备腹水抗体;采用间接ELISA法鉴定抗体亚型和测定抗体效价。结果得到6株能稳定分泌单克隆抗体的杂交瘤细胞株;抗体经鉴定均为IgG1、κ型,效价均达10-5以上。结论所获得的6株杂交瘤细胞株均有较强稳定分泌抗-hCG单克隆抗体的能力。这为有关hCG的检测、hCG本身相关研究以及避孕疫苗的研制打下了基础。

Objective To prepare monoclonal antibody against hCG. Methods Balb/c mice were immunized with hCG protein, and spleen cells from the mice were fused with myeloma cells from isogenic mice at ratio of 8∶1. Positive clones were screened by indirect enzyme-linked immunoadsorbent assay (ELISA), and then the clones were subcloned with limiting dilution and amplified further. After intraperitoneal injection of hybridoma cells, mouse ascites antibody was prepared. Isoforms, titres, specificity, and cross-reactivity of the ascites antibody were identified and determined by indirect ELISA. Results Six strains of hybridoma cell lines were obtained, which could secret MAb stably. Prepared ascites MAb from all mice were identified as IgG1 and κ type with titre of more than 10^(-5). Conclusion The obtained 6 strains of hybridoma cell lines can secret MAb stably. This research benefits hCG detection, correlative study of hCG, and preparation of contraceptive vaccines.