CAJ | 학술논문

目的制备人绒促性素(hCG)单克隆抗体.方法 hCG抗原免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/O按5:1融合,间接ELISA法筛选阳性孔,有限稀释法进行克隆化培养;制备腹水抗体;间接ELISA法测定抗体效价:采用HiTrap rProtein A FF亲和色谱柱纯化抗体.结果得到2株能稳定分泌单克隆抗体的杂交瘤细胞株,细胞培养上清中抗体效价达10~(-3)以上,腹水抗体效价达10~(-7)以上,纯化后的单抗纯度迭98%以上,回收率达75%.结论成功获得两株能稳定分泌单克隆抗体的杂交瘤细胞,可用于早孕、肿瘤等诊断的研究.
목적 제비인융촉성소(hCG)단극륭항체.방법 hCG항원면역BALB/c소서,취기비세포여소서골수류세포SP2/O안5:1융합,간접ELISA법사선양성공,유한희석법진행극륭화배양;제비복수항체;간접ELISA법측정항체효개:채용HiTrap rProtein A FF친화색보주순화항체.결과 득도2주능은정분비단극륭항체적잡교류세포주,세포배양상청중항체효개체10~(-3)이상,복수항체효개체10~(-7)이상,순화후적단항순도질98%이상,회수솔체75%.결론 성공획득량주능은정분비단극륭항체적잡교류세포,가용우조잉、종류등진단적연구.
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 .