高效疏水色谱法对还原变性核糖核酸酶的折叠研究

Refolding of Reduced and Denatured Ribonuclease A by High Performance Hydrophobic Interaction Chromatography

用高效疏水色谱(HPHIC)研究了还原变性核糖核酸酶(RNase A)在HIC上流动相中脲浓度、流动相流速及蛋白浓度对还原变性RNase A复性的影响.发现还原变性RNase A在脲浓度为2.0mol/L时,复性效率可达80%以上.在蛋白浓度为20 mg/mL时,复性效率可达86.4%,当蛋白浓度过高时,在HIC上因聚集严重导致复性效率降低.流动相的流速对复性影响不大.

Reduced and denatured RNase A by using high performance hydrophobic interaction chromatography （HPHIC） was studied. The effects of urea concentration, flow rate of mobile phase and protein concentration were also investigated. When the urea concentration was 2.0 mol/L, the bioactivity recovery of reduced and denatured RNase A was more than 80%. When the protein concentration was 20 mg/mL, its bioactivity recovery was 86.4 %. When the protein concentration was much higher, its bioactivity would be highly reduced because of its great mass. Its flow rate of mobile phase had little effect on its bioactivity recovery.