重组猪α干扰素的纯化及其对猪繁殖与呼吸综合征病毒的抑制作用

Purification of recombinant porcine interferon-α and its inhibitory effect on porcine reproductive and respiratory syndrome virus

建立重组猪α干扰素（rPoIFN-α）的毕赤酵母分泌表达工艺,甲醇诱导84 h时,rPoIFN-α的表达量最高,达650.3 g/mL。随后,依次采用亲和、离子交换、分子筛层析对获得的rPoIFN-α进行纯化,得到纯度为95%的rPoIFN-α,生物活性为5.63×106IU/mg。细胞病变抑制结果表明,以341 IU/mL的rPoIFN-α预处理Marc145,可以完全抑制猪繁殖与呼吸综合征病毒（PRRSV）的增殖;在PRRSV感染后的24 h,1 367IU/mL的rPoIFN-α可以完全抑制PRRSV的增殖。结果表明,获得的rPoIFN-α可有效抑制PRRSV的增殖,为rPoIFN-α的临床应用奠定了科研基础。

The secretory expression of recombinant porcine interferon-α（rPoIFN-α） in Pichia pastoris was established.The highest concentration of rPoIFN-α reached 650.3 g/mL at 84 h after initiating induction.The purity of rPoIFN-α from using affinity chromatography,ion exchange column chromatography and gel filtration column chromatography in turn was 95% and the inhibitory activity of purified rPoIFN-α against vesicular stomatitis virus was about 5.63 ×106 IU/mg using cytopathogenic effect inhibition assay.341 IU/mL rPoIFN-α pretreatment of Marc145 cells could completely inhibit the proliferation of porcine reproductive and respiratory syndrome virus（PRRSV）.The treatment of PRRSV infected cells with 1 367 IU/mL rPo IFN-α could completely inhibit the proliferation of PRRSV.The results indicated that rPoIFN-α has inhibitory effects on PRRSV infection in vitro,which provided scientific foundation for clinical application of rPoIFN-α.