Simple and rapid purification of monoclonal antibodies from cell culture supernatants and ascites fluids by hydroxylapatite chromatography on analytical and preparative scales.
Simple and rapid purification of monoclonal antibodies from cell culture supernatants and ascites fluids by hydroxylapatite chromatography on analytical and preparative scales.
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通过羟基磷灰石色谱法在分析和制备规模上从细胞培养上清液和腹水中简单快速地纯化单克隆抗体。
DOI：
10.1089/hyb.1987.6.219
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Kennett,RH
中科院分区：
文献类型：
--
作者：
Bukovsky,J;Kennett,RH
本文描述了一种从腹水和细胞培养上清液中纯化小鼠和人单克隆抗体的简单快速方法。该方法基于羟基磷灰石（HAP）柱层析，适用于分析级和制备级。在我们纯化单克隆抗体的工作中，我们发现杂质的单步洗脱和抗体的线性梯度洗脱相结合，可以很好地从细胞培养和腹水中纯化抗体。该程序在高流速下提供非常好的分辨率。将细胞培养上清液以2-3ml/min的速率泵送到制备柱上，无任何可测的背压。这种结合与流速无关。该方法已成功地从细胞培养上清液中纯化了几种不同亚型的单克隆抗体。
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 2-3ml/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.