【摘要】 利用稀释法、凝胶过滤、脲浓度梯度凝胶过滤 3种方法研究了抗肿瘤血管抗体功能片段VH/κ的体外复性 .通过考察复性液中还原型谷胱甘肽 /氧化型谷胱甘肽 (GSH/GSSG)比例、精氨酸浓度、pH值、洗脱速度、变性液中蛋白质浓度、凝胶过滤脲梯度长度等因素 ,发展了脲梯度凝胶过滤柱复性VH/κ的方法 .结果表明 ,与传统的稀释法和柱复性法相比 ,脲梯度法复性获得的VH/κ的活性回收率和相对亲和力均有显著提高 .更多还原

【Abstract】 The functional fragment V H/κ of an anti tumor antibody AA98 was refolded in vitro by dilution refolding, gel filtration chromatography and urea gradient gel filtration, respectively. Concerning various factors in the renaturing buffer, such as GSH/GSSG ratio, arginine concentration, pH, flow rate, protein concentration in denaturing buffer and linear gradient, a urea gradient gel filtration method for V H/κ refolding was established. As a result, compared with the traditional dilution refolding and gel filtration, the activity recovery of V H/κ obtained by urea gradient gel filtration was significantly improved.更多还原