摘要
目的: 研究变性蛋白质在高效疏水作用色谱(HPH IC)上的保留行为及复性效率. 方法: 分别用盐酸胍(Gu HCl)、尿素(Urea)和十二烷基磺酸钠(SDS)将核糖核酸酶(RNase)等10种标准蛋白质变性,然后在流动相为硫酸铵的条件下,用HPHIC对变性蛋白质进行复性,并测定其保留时间、Z值、质量及活性回收率. 结果: 在10种变性蛋白质中有5种在HPHIC上得到复性,其保留时间、峰形、Z值与天然蛋白基本一致,其中溶菌酶和核糖核酸酶的质量和活性回收率高,复性效果好. 结论: 用HPHIC对变性的标准蛋白质进行复性,得到较好的复性效果.
AIM: To investigate the refolding of the denatured proteins by high performance hydrophobic interaction chromatography(HPHIC).METHODS: Ten standard proteins were denatured by guanidine hydrochloride and urea or sodium dodecyl sulfate,and then were refolded by HPHIC in the mobile phrase of (NH 4) 2SO 4.The retention time, Z value, mass and bioactivity recovery were detected. RESULTS : Among the ten denatured proteins,five proteins were refolded by HPHIC,in which lysozyme and ribonuclease had high mass and bioactivity recovery and their retention time,peak form and Z value were identical with those of the respective natural proteins.CONCLUSION: HPHIC has the advantage of good renaturation efficiency.
出处
《第四军医大学学报》
北大核心
2005年第8期682-684,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30080007)
关键词
蛋白质变性
蛋白质复性
高效疏水作用色谱
protein denaturation
protein renaturation
high performance hydrophobic interaction chromato graphy
