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抗肿瘤血管抗体AA98功能片段V_H/κ的脲浓度梯度柱复性
Expression and Refolding of an Anti-tumor Antibody AA98 Fragment V_H/κ
【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 .
【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.
- 【文献出处】 生物化学与生物物理进展 ,Progress In Biochemistry and Biophysics , 编辑部邮箱 ,2002年04期
- 【分类号】Q51
- 【被引频次】3
- 【下载频次】81