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用适合杂交瘤生长的基础培养基对血清进行错流渗滤、灭菌和蛋白质减量。

Cross flow diafiltration of serum with basal medium suitable for growth of hybridomas, sterilization and protein reduction.

作者信息

Jungbauer A, Wenisch E, Steindl F, Himmler G, Reiter S, Rüker F, Wagner K, Katinger H

出版信息

Dev Biol Stand. 1987;66:87-90.

PMID:3582774
Abstract

Fetal Calf Serum (FCS) was extensively extracted by cross flow diafiltration (Pellicon, Millipore) and sterilized using the basal growth medium (DMEM) for the extraction. Ultrafiltration membranes of 10(5) and 3 X 10(5) Dalton cut off were used respectively. The diafiltrates were used for hybridoma cultivation and the results of growth and mAb-production were compared with standard medium (DMEM + 5% FCS). Slightly reduced mAb-titers were achieved. These were, however, compensated by decreased concentration of contaminating protein and higher specific mAb/protein ratio as examined by SDS-PAGE and enzyme linked immuno electro transfer blot (EITB).

摘要

胎牛血清(FCS)通过错流渗滤(Pellicon,密理博公司)进行广泛提取,并使用基础生长培养基(DMEM)进行提取过程中的灭菌处理。分别使用截留分子量为10(5)和3×10(5)道尔顿的超滤膜。渗滤液用于杂交瘤培养,并将生长和单克隆抗体产生的结果与标准培养基(DMEM + 5% FCS)进行比较。单克隆抗体滴度略有降低。然而,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和酶联免疫电转印(EITB)检测发现,污染蛋白浓度降低以及单克隆抗体/蛋白比例提高,弥补了滴度的下降。

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