层析介质Capto Core400和Sephacryl S-1000纯化乙型肝炎病毒核心蛋白病毒样颗粒效果的比较

Purification of virus-like particles of hepatitis B virus core protein by Capto Core400 and Sephacryl S-1000 as chromatographic media

目的比较层析介质Capto Core400和Sephacryl S-1000纯化乙型肝炎病毒核心蛋白(hepatitis B virus core protein,HBc)病毒样颗粒(virus-like particles,VLPs)的效果。方法采用两种层析介质纯化相同3批HBc VLPs蛋白粗提液,BCA法检测纯化样品的蛋白浓度,计算蛋白回收率;体积排阻色谱法检测纯度;马尔文粒径仪检测HBc VLPs粒径;透射电子显微镜观察HBc VLPs形态。结果层析介质Sephacryl S-1000和Capto Core400纯化HBc VLPs的蛋白回收率分别为75.5%和70.5%,蛋白纯度分别为87%和99%,粒径大小均为(30±4)nm,均一性良好;与Sephacryl S-1000比较,经Capto Core400纯化的HBc VLPs分布更均匀。结论Capto Core400纯化HBc VLPs的纯度及颗粒分布均匀性更佳,且该层析介质装量小,纯化时间短,更适用于HBc VLPs的规模化生产。

Objective To compare the effects of Capto Core400 and Sephacryl S-1000 as chromatogrphic media in purification of virus-like particles(VLPs)of hepatitis B virus core protein(HBc).Methods Three batches of crude protein extracts of HBc VLPs were purified by the two kinds of chromatography media separately,and determined for protein concentration,based on which the protein recovery rate was calculated.The purity of purified protein was determined by size exclusion chromatography,while the size of HBc VLPs by Malvern particle sizer,and the morphology was observed by transmission electron microscopy.Results The recovery rates of HBc VLPs purified with Sephacryl S-1000 and Capto Core400 were 75.5%and 70.5%,while the purities were 87%and 99%,respectively.However,both the sizes of HBc VLPs purified by the two kinds of media were(30±4)nm,indicating a good homogeneity.Compared with that by Sephacryl S-1000,the HBc VLPs purified by Capto Core400 were distributed evenly.Conclusion The HBc VLPs purified by Capto Core400 showed high purity and were distributed more evenly,which was small in filling quantity and short in purification time,and was suitable for large-scale production of HBc VLPs.