论文部分内容阅读
制备抗促乳素(prolactin,PRL)的单克隆抗体。应用PRL纯品免疫BALB/c小鼠,被免疫小鼠的脾细胞与SP2/0骨髓瘤细胞融合,间接ELISA筛选阳性克隆,将阳性克隆多次有限稀释得到抗PRL阳性单克隆细胞株,杂交瘤细胞上清分离法和诱生腹水法制备单克隆抗体,测其效价。共建立5株持续分泌抗体的杂交瘤细胞,分别命名为AA5B1、AB6C8、BB3C1、CA6E4、CE2H6。杂交瘤细胞株冻存4个月后复苏培养,形态良好,生长旺盛。培养液上清及腹水效价的OD值略有升高,说明杂交瘤细胞株稳定。5株杂交瘤细胞染色体众数均为99～105,并可见到中部和亚中部着丝点的骨髓瘤细胞标志染色体,说明5株杂交瘤细胞确为SP2/0骨髓瘤细胞与被免疫小鼠的脾细胞融合而产生的。
Preparation of anti-prolactin (prolactin, PRL) monoclonal antibody. BALB / c mice were immunized with purified PRL. The spleen cells of immunized mice were fused with SP2 / 0 myeloma cells. The positive clones were screened by indirect ELISA. The positive clones were diluted for several times to obtain anti-PRL positive monoclonal cell lines. Tumor cell supernatant separation and ascites induced monoclonal antibody preparation, measuring the titer. A total of five hybridomas secreting antibodies were established and named AA5B1, AB6C8, BB3C1, CA6E4 and CE2H6, respectively. The hybridoma cell lines were resuscitated and cultured 4 months after cryopreservation, with good morphology and vigorous growth. OD value of culture supernatant and ascites titer increased slightly, indicating that the hybridoma cell line is stable. The chromosome numbers of the 5 hybridoma cells were all from 99 to 105, and the chromosomal markers of myeloma cells in the centromere and in the middle of the centromere were observed, indicating that the 5 hybridoma cells were indeed SP2 / 0 myeloma cells and the immunized mice Of spleen cells produced by fusion.