摘要
用人甲胎蛋白(AFP)免疫BALB/c小鼠的脾细胞与SP2/O骨髓瘤细胞融合,获得两株分泌抗人AFP单克隆抗体(McAb)的杂交瘤细胞株(G2和G10)。用酶联免疫吸附试验(ELISA)检测细胞培养上清液和小鼠腹水的特异性抗体效价,分别为2～4×10^(-3)和10^(-6)～10^(-7)。两株杂交瘤细胞染色体的众数为104～108条。经体外连续传代及冻存半年复苏后,仍具有分泌抗体能力,说明其性能稳定。两株所分泌的抗体,均属IgGl型。
This paper reported the establishment of two hybridoma cell lines secreting anti-human AFP monoclonal antibodies,by using the method of fusing the mouse myeloma cell SP2/O and the spleen cell of BALB/c mice immunized by purified human AFP.These two hybridoma cell lines were named G2 and G10.Hybrid clones producing specific antibody were screened by enzyme-linked immunosorbent assay(ELISA)techniques.The monoclonal antibodies titer in the supernatant of the cultured hybrid cells and in the ascitic fluid of the mouse ranged from 2～4×10^(-3)and 10^(-6)～10^(-7) respectively.G2 and G10 were stable in secreting anti-human AFP McAb in the tissue culture for over four months,although they had been stored in liquid nitrogen for over eight months.Karyotype analysis of the hybridoma cell G2 and G10 showed that their chromosome number were 104 and 108 respectively.These McAb belonged to the mouse IgGl subtype,according to the analysis using double diffusion precipitation.
