白细胞介素6单克隆抗体的研制

THE PREPARATION AND PRELIMINARY DETERMINATION OF ANTI IL 6 MONOCLONAL ANTIBODY

采用纯化的工程菌表达产物白细胞介素６（Ｉｎｔｅｒｌｅｕｋｉｎ６，ＩＬ－６）免疫ＢＡＬＢ／Ｃ小鼠，取小鼠脾细胞与骨髓瘤细胞（ＳＰ２／０）在ＰＥＧ作用下进行融合，通过ＥＬＩＳＡ筛选并多次亚克隆，获得了１株能分泌抗ＩＬ－６单克隆抗体的杂交瘤细胞Ｉｃ－１．３．４．５。Ｗｅｓｔｅｒｎｂｌｏｔ显示：抗体结合抗原的分子量与ＩＬ－６相符，为特异性抗ＩＬ－６抗体。亚类鉴定表明：该细胞分泌的单克隆抗体为ＩｇＧ１亚类，ｋａｐｐａ型。染色体数目为１００条左右。腹水效价为１１０４，无血清培养液效价为１１０２。此杂交瘤细胞株体外传代培养６个月以上，ＥＬＩＳＡ检测抗体滴度无明显降低。

Balb/c mice were immunized with interleukin 6(IL 6)antigen purified from engineered E.coli strain,their spleen cells were fused with myeloma cells by polyethyleneglyco(PEG).After making ELISA screening and subcloning we obtained one positive hybridoma cell strain Ic 1.3.4.5.Western blot demonstrated that antibody was peculiar to combine with IL 6 antigen.The monoclonal antibody was IgG1 subclass,kappa type.Chromosomal numbers were 100 or so.We cultivated and frozen cells,prepared ascetic fluid and serum free supernatant.The titer of ascetic fluid was 1∶10 4 and serum free supernatant was 1∶10 2.The hybridoma cell was bequeathed more than six months and the titer of antibody had not obviously changed.