猪α-干扰素在毕赤酵母中的分泌表达和下游工艺的研究

The high serection and down-stream technology of porcine interferon-α in the yeast Pichia pastoris

通过小量发酵、摇瓶发酵、大量发酵,研究猪α-IFN在毕赤酵母中表达的下游工艺,微量细胞板法测猪α-IFN体外抗VSV、PRRSV、TGEV的活性。猪α-IFN抗VSV的活性达到4.04×10^6I U/L,在PK15上抗TGEV的活性为10^7-10^8I U/L,在Marc 145上抗PRRSV的活性为10^6-10^7I U/L。30℃诱导培养96 h、甲醇添加1%、pH值6.0、DO20%、0.25%-0.4%甲醛灭活,获得高活性的、安全的IFN。仔猪免疫后临床症状正常,未出现病理学变化。猪α-IFN体外抗病毒活性高,发酵后的抗病毒活性未下降,免疫动物未产生临床症状和病理变化,可用于临床治疗病毒性疾病。

Study the down-stream technology,such as the antiviral activities of porcine interferon-α,fermentation,toxicology and safety pharmacology.The antiviral activities of porcine interferon-α were observed on the 96-wellmicroplate and the fermentation in test-tube,in shake flasks and fermentor.The antiviral activities of porcine IFN-α on VSV,TGEV and PRRSV were investigated in Vero,PK15 and Marc145,and their activities were 1.370×10^9 IU/L,10^8-10^9 IU/L and 10^8-10^9 IU/L.The α-IFN was inactivated with 30℃,96 h,1% methanol,pH 6.0,DO 20%,0.25%-0.4% formaldehyde.There were on clinical symptoms and meaningful histologic alterations in 44 days old pigs after immunization.