Construction and expression of soluble and functional Recombinant Porcine Interferon alpha in Escherichia coil
Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers.Recombinant IFNs have been used as anti-infectious agents exhibiting a broad range of antiviral and immunomodulatory properties in both human and domestic animals.Escherichia coli is the most extensively used host for the production of recombinant proteins.However,most of the eukaryotic proteins are typically obtained as insoluble,misfolded inclusion bodies that need solubilization and refolding.In this report,recombinant porcine interferon alpha was expressed in Escherichia coli as soluble form,and was purified to homogeneity using essentially two-step chromatographic procedures,i.e.immobilized metal-ion-affinity chromatography and DEAE anion exchange chromatography.The antiviral activity of the final rPoIFNα protein was 1.1x 106 IU/ ml protein,which is comparable with the HuIFNα1 standard.To establish methods and requirements for quality control of rPoIFNα protein,the research of quality control includes biological activity (anti-virus activity),purity (included electrophoresis assay and HPLC assay ),protein content,molecular weight,isoelectric point,test for bacterial endotoxin,Ultraviolet spectroscopy,peptide mapping,N-terminal amino acid sequence.
赵俊 王明丽 沈咏舟 张俊玲 王兴满 李京培 徐敬东 韩国祥
安徽医科大学微生物学教研室;安徽九川生物科技有限公司 安徽九川生物科技有限公司
国内会议
第五届中国兽药大会(中国畜牧兽医学会动物药品学分会2014年学术年会、中国畜牧兽医学会生物制药学分会暨中国微生物学会兽医微生物学专业委员会联合学术论坛)
沈阳
英文
270-279
2014-09-13（万方平台首次上网日期，不代表论文的发表时间）