鼠源性抗人IgMμ链单克隆抗体的制备和鉴定

目的 :建立能稳定分泌高效价抗人 Ig Mμ链单克隆抗体的细胞株 ,收获并纯化抗体用于进一步研究。方法 :小鼠免疫脾细胞与骨髓瘤细胞用 PEG融合 ,小鼠体内诱生腹水的方法收获大量单克隆抗体 ,辛酸硫酸铵提纯 ,并鉴定其纯度。结果 :获得 3个稳定分泌抗体的细胞株 M1、M2、M3,鉴定其亚类分别为 Ig G3、Ig G2 a、Ig G1;腹水抗体纯化后获得纯化抗体含量分别为 0 .36 mg/ ml、0 .2 8mg/ ml、3.71mg/ ml。结论 :用 PEG进行细胞融合 ,辛酸硫酸铵盐析法纯化能够获得较高效价和纯度的抗体

Generation of Mouse Monoclonal Antibodies Against Human IgM μ Chains

Objective To establish cell lines that c ould stably secrete specific mAbs,and to generate purified mAbs for next researc h.Methods The splenocytes immunized with human IgM were fused wit h myeloma cells SP2/0. Ascitic mAbs collected from mice inoculated with hybridom a cells were purified by method of caprylic acid-ammonium sulfateprecipitation. Results Three hybridomas whose subtypes were IgG3, IgG2a and IgG w ere obtained.The concertaions of purified mAb s were 0.36 mg/ml, 0.28 mg/ml a nd 3.71 mg/ml.Conclusion It is proper to use PEG and caprylicacid- ammonium sulfate precipitation to get excellent mAbs.