Witt S, Ziegler B, Blumentritt C, Schlosser M, Ziegler M
Institut für Diabetes, Gerhardt Katsch, Karlsburg.
Allerg Immunol (Leipz). 1991;37(2):67-74.
A simple procedure for the production of pure monoclonal antibodies (mab) in dialysis tubing has been used. Hybridomas which produce pancreatic islet cell reactive monoclonal antibodies were grown in dialysis tubing containing serum free medium. The dialysis tubing was inserted into a flask with medium containing 7.5% foetal calf serum. The flask was placed on a roller and medium was changed every two days. The optimal time for harvesting the mab could be shown to be after 10 days of culture with a 4 fold increased immunoglobulin concentration in comparison to a conventional hybridoma culture. Immunoglobulin concentrations up to 110 mg/l and cell yields of 1.3 x 10(6)/ml have been obtained. The low concentration of contaminating low molecular weight proteins in the supernatant facilitated or saved purification of mab.
已采用一种在透析袋中生产纯单克隆抗体(mab)的简单方法。产生胰腺胰岛细胞反应性单克隆抗体的杂交瘤细胞在含有无血清培养基的透析袋中培养。将透析袋插入装有含7.5%胎牛血清培养基的烧瓶中。烧瓶置于滚轴上,每两天更换一次培养基。结果表明,收获单克隆抗体的最佳时间是培养10天后,与传统杂交瘤培养相比,免疫球蛋白浓度提高了4倍。已获得高达110mg/l的免疫球蛋白浓度和1.3×10(6)/ml的细胞产量。上清液中低分子量污染蛋白的低浓度便于或节省了单克隆抗体的纯化。