摘要
高效表达了ＨＣＶ核心区基因抗原之后，对表达蛋白Ｃ_（２７）进行了纯化。经研究，重组蛋白是以包涵体形式存在于宿主菌内的。Ｃ_（２７）重组蛋白分别经过包涵体洗涤、ＤＥＡＥ阴离子交换层析和Ｓ－２００分子筛两步柱层析纯化之后，纯度大于９５％，纯化得率为５３．２％，总回收率为１７．９％。纯化工艺流程简单、得率高，适合向规模化生产发展。
The expressed HCV core recombinant fusion protein C_(27) was verified to be in the form of inclusion bodies(IBs)in E. coli 06 by the methods of ELISA,SDS-PAGE and electron micrograph. The cells were collected by centrifugation,then broken with ultrasonication,repeated freezing and thawing. After being washed several times,the C_(27) in IBs protein is 51%. Then the washed IBs was solibized in buffer A (50 mmol/L Tris Cl,8 mol/L urea, PH8. 5)and applied on the DEAE-Sepharose-FF column equilibrated with the same buffer. The column was washed with a linear NaCl concentration.After DEAE purification, the purity of C_(27)increased to·88. 3%.The purified protein from DEAE was applied on a S-200 column(3 cm ×90cm). After this procedure,the C_(27)purity was more than 95% from analysis by SDS-PAGE and staining with Coomssie Blue.
