Yurov G K, Neugodova G L, Verkhovsky O A, Naroditsky B S
All-Russia Research Institute of Agricultural Biotechnology, Moscow.
J Immunol Methods. 1994 Dec 28;177(1-2):29-33. doi: 10.1016/0022-1759(94)90140-6.
A thiophilic adsorption method has been developed for rapid purification and separation of mouse F(ab)2 and Fc fragments obtained after proteolytic digestion of IgG1 monoclonal antibodies. Partially purified Mabs were digested with papain. Thiophilic chromatography was performed using stepwise elution with decreasing concentrations of ammonium sulphate. Most contaminating proteins did not react with the thiophilic adsorbent, and chromatography efficiently resolved the F(ab)2 and Fc fragments, as judged by electrophoresis. Fractions containing the F(ab)2 fragments retained about 90% of the total antibody activity loaded onto the column.
已开发出一种嗜硫吸附方法,用于快速纯化和分离IgG1单克隆抗体经蛋白水解消化后获得的小鼠F(ab)2和Fc片段。将部分纯化的单克隆抗体用木瓜蛋白酶消化。采用硫酸铵浓度递减的分步洗脱进行嗜硫色谱分析。大多数污染蛋白不与嗜硫吸附剂反应,通过电泳判断,色谱法能有效分离F(ab)2和Fc片段。含有F(ab)2片段的馏分保留了加载到柱上的总抗体活性的约90%。