重组人γ-干扰素单克隆抗体杂交瘤细胞建株研究

本文报道重组人γ-干扰素单克隆抗体杂交瘤细胞株建立的方法及结果。为获得供融合用的小鼠脾细胞,用两种不同的方案免疫了两组(每组3只)BALB/c小鼠。血清抗体效价最高的第4号小鼠,于加强免疫4天后取脾,制成脾细胞悬液。按照常规方法与小鼠骨髓瘤SP 2/0·Ag 14细胞融合。采用直接中和试验法筛选融合细胞孔的上清液,从中选出两株(A3和B3)分泌抗重组人γ-干扰素的杂交瘤细胞。交叉中和试验显示,A_3和B_3抗体只中和重组或自然的人γ-干扰素;与α-干扰素或β-干扰素无反应。此两株单克隆抗体的效价分别超过128,000和25,600。双扩散试验证明,A_3和B_3均为小鼠IgG_1型抗体;液氮中保存半年以上,仍保持原有的各种特性。

ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST HUMAN RECOMBINANT INTERFERON-GAMMA

This paper reports the methods and results of establishment of hybridoma cell lines secreting monoclonal antibodies (McAb) against human recombinant interferon-gamma (Hu rIFN-gamma). To gain immunized mouse spleen cells for hybridization, two groups of BALB/c mice (three mice each group) were immunized in two different projects. Mouse No. 4 with the highest serum antibody titer was used for fusion 4 days after boosting im-munization. The spleen cell suspension was made and fused with mouse myeloma cell line SP 2/0·2Ag 14 based on the routine procedure. Culture supernatants in the wells containing hybridoma cells were screened by direct neutralization test. Two hybridoma cell lines, named A3 and B3, secreting McAb toHu rIFN-gamma were selected out. The results of crossing neutral ization test showed that the supernatants from A3 and B3 could only neutralize Hu rIFN-gamma and natural interferon-gamma,but not IFN-alpha and IFN-beta. The antibody liters of the two hybridomas were more than 128,000 and 25,600, respect-ively. The double diffusion test showed that both A3 and B3 supernatants contained mouse IgG 1. After storage in liquid nitrogen for six months, the hydridomas A3 and B3 still retained their original characteristics.