Bukovsky J, Kennett R H
Hybridoma. 1987 Apr;6(2):219-28. doi: 10.1089/hyb.1987.6.219.
A simple and rapid method for the purification of murine and human monoclonal antibodies from ascites fluids and cell culture supernatants is described. The method, based on the use of hydroxylapatite (HAP) column chromatography, is applicable on both analytical and preparative scales. In our work on purification of monoclonal antibodies, we have found that the combination of a single step elution of impurities followed by linear gradient elution of antibody provides an excellent purification of the antibody from cell culture and ascites fluids. The procedure provides very good resolution at high flow rates. The cell culture supernatant can be pumped on the preparative column at the rate of 2-3 ml/min without any measureable back pressure. The binding is independent of the flow rate. This method has been successfully used to purify several monoclonal antibodies of different subtypes from cell culture supernatants.
本文描述了一种从腹水和细胞培养上清液中纯化鼠源和人源单克隆抗体的简单快速方法。该方法基于使用羟基磷灰石（HAP）柱色谱，适用于分析和制备规模。在我们纯化单克隆抗体的工作中，我们发现先进行一步杂质洗脱，然后进行抗体线性梯度洗脱，可从细胞培养物和腹水中出色地纯化抗体。该程序在高流速下具有很好的分辨率。细胞培养上清液可以以2 - 3 ml/分钟的流速泵入制备柱，而没有任何可测量的背压。结合与流速无关。该方法已成功用于从细胞培养上清液中纯化几种不同亚型的单克隆抗体。
