摘要
作者采用小鼠骨髓瘤细胞系与破伤风类毒素免疫的人外周血淋巴细胞进行融合,对影响人-鼠杂交瘤细胞制备人单抗成功的因素进行了探讨。按免疫时间(天)及体外刺激与否分为四组、其中以 PWM+TT 体外刺激 PBL(比体外不刺激)的细胞融合率高,且易建成稳定分泌抗体的细胞系。经5次克隆化后获得5株抗 TT 的人单抗杂交瘤细胞系。经双扩确定 IgG2株,IgM3株。冻存10个月后复苏,传代及适当的克隆化后仍为阳性。上清液中人 IgM 的分泌量为0.42～1.15μg/ml。经染色体鉴定确为人-鼠杂种细胞。
The cells from mouse myeloma cell line SP2/0 were fused with human peripheral blood lymphocytes from the donors primed with a conventional dosage of tetanus toxoid (TT).Four experimental groups were divided by immunization time and presence or absence of stimulation in vitro.The group with pokeweed mitogen plus TT stimulation in vitro has the highest fusion rate and easily establishes cell line secreting antibodies stably.Five hybridoma cell lines secreting human monoclonal antibodies against tetanus toxoid were obtained after clonization for five times Double diffusion assay proves that two of the ceil lines secret IgG and three secret IgM.They produced antibodies against TT after freezing for 10 months,revivification,generation and clonization. Karyotype analysis confirmes that they are human-mouse hybridomas.
出处
《南京铁道医学院学报》
1991年第4期210-213,共4页
Journal of Nanjing Railway Medical College
基金
江苏省卫生厅科研基金
