分泌抗白喉类毒秦单克隆抗体杂交瘤细胞株的建立

我们用白喉类毒素免疫小鼠的脾细胞与NS-1小鼠骨髓瘤细胞缸合,获得一株分泌抗白喉类毒素单克隆抗体的杂交瘤细胞株。检测此杂交瘤细胞培养上清液中特异性抗体效价:间接血凝法为1:640～1280,ELISA法为1:800～1600。细胞注入同系小鼠可诱生含高效价抗体的腹水(高于培养液1000倍)。用免疫双扩散法鉴定此单克隆抗体属小鼠I。GI亚类。杂交瘤细胞染色体数为86～102。此细胞株经连续传代六个月和在液氮中冻存复苏后仍保持稳定的分泌抗体能力。

Establishment of hybridoma cell line secreting monoclonal antibody against diphtheria toxoid

Spleen cells taken from BALB/c mice Immunized with diphtheria toxoid were fused with NS-1 myeloma cells in the presence of 50%PEG (M,W4,000). On the 6th day hybridoma cells were found, with a fusing rate of about 73.8%. Two hybridoma cell lines secreting specific antidiphtheria toxoid antibodies (4D8, 5D10) were selected by means of ELISA and passive hemagglutination (PHA). In the course of cloning, one of them (4D8) lose its ability, while the other (5D10) kept its ability to secrete McAb.The titer of McAb produced by 5D10 varied with the sources of the test samples, e.g. 1 :640- 1 :1280 by PHA and 1 :800-1:1600 by ELISA in cell culture supernatant 1 :106-1 :107 by ELISA and PHA in ascites.McAb were identified as mouse IgG1 with standard rabbit anti-mouse antiserum.The numbers of chromosomes were between 86-102. When stored in liquid nitrogen for 6 months supplemented with calf serum containing 10% dimethyl sulphoxide, the hybridoma cells maintained their ability to secrete specific McAb.