肿瘤坏死因子α杂交瘤细胞系的建立

ESTABLISHMENT OF THE HYBRIDOMA CELL STRAINS SECRETING ANTI-TUMOR NECROSIS FACTOR MONOCLONAL ANTIBODY

应用细胞融合技术建立了2株(3A_9、4B_7)能稳定分泌肿瘤坏死因子单克隆抗体(TNFMcAb)的杂交瘤细胞系,经鉴定:(1)二株均为IgG1;(2)染色体为101条;(3)二株McAb腹水1:20稀释与10U的TNFa进行免疫沉淀反应,结果TNF几乎全部失去活性(将近90％以上活性丧失);(4)效价测定,腹水为1×10^((-4)~2)×10(-4),培养上清液为5×10^((-2)~1)×10^(-3)或更高;(5)TNF McAb置4℃2个月,室温(25℃),10d抗体效价基本稳定;(6)杂交瘤细胞在液氮中冻贮2个月后复苏,细胞生长良好,抗体分泌正常。本研究为rHTNFa的纯化、生物学特性研究、治疗感染性内毒素休克、研制TNF检测药盒提供了物质基础。为进一步研究TNF与重型肝炎的致病机理、筛选治疗重型肝炎药物、判断病情预后有十分重要的价值。

In our laboratory, we have established 2 strains of hybridoma celIs(3Ag,4B7), which can stably secrete anti-tumor necrosis factor antibodies (anti-TNF McAb). We found that the two strains possess the following characteristics: (1) the secreted antibodies were detected to be IgG 1 type; (2) its chromosome number was found to be 101; (3) when the ascitic fluid secreted by the monoclonal antibodies was used in the dilution of 1:20 to act upon 10U of TNFa,over 90% of TNFa lost its biological activity; (4) the efficacy of the ascitic fluid was found to be(1~2)×10~4, while that of the culture supernatant to be 5×10-2~1×10~3, or higher; (5) the liter of the secreted anti-TNFMcAb is stable, after 2 months of storage at 4℃, or 10 days at 25℃, or 3712; (6)the hybridoma cells grew well with its normal McAb secretion after being thawed from eryopreservation after the 2 months in liquid nitrogen for a period of 2 months.