重组猪α干扰素理化性状及其体外抗病毒活性研究  

Study on the Physicochemical Characters of rPoIFN-α and Its Antiviral Activity in vitro

作　　者：汤仁树[1] 赵俊[1] 王明丽[1] 

机构地区：[1]安徽医科大学微生物学教研室教育部"重要遗传疾病基因资源利用"重点实验室省部共建,安徽合肥230032

出　　处：《安徽农业科学》2010年第8期3928-3930,3934,共4页Journal of Anhui Agricultural Sciences

基　　金：安徽省教育厅自然科学研究计划项目(2004kj218);安徽省科技攻关计划重大科技专项(08010302179)

摘　　要：[目的]研究重组猪α干扰素(rPoIFN-α)半成品理化性状并对其体外抗病毒活性进行测试与鉴定。[方法]用HEp-2/VSV体系对3批rPoIFN-α蛋白进行抗病毒活性检测,以重组人IFN-α为参比品,测定干扰素效价;用0.25%胰蛋白酶HCl以及鼠抗猪α干扰素单克隆抗体作用已知效价的rPoIFN-α半成品,并检测各批次抗病毒活性,对rPoIFN-α理化性状进行评价;在猪肾细胞株(PK-15)上检测rPoIFN-α对猪细小病毒(PPV)和猪伪狂犬病毒(PRV)的致细胞病变抑制效应,观察rPoIFN-α的体外抗病毒活性。[结果]HEp-2/VSV体系滴定rPoIFN-α半成品效价可达1.5×105IU/ml,比活性达1.1×106IU/mg;rPoIFN-α经0.25%胰蛋白酶37℃作用1h,效价残留率低于1%,经HCl(pH=2.0)处理72h效价残留率高达95%以上,经56℃处理30min效价残留率高于47%,经鼠抗猪α干扰素单克隆抗体37℃作用1h后效价残留率约为1%;体外抗病毒试验表明,用50和500IU/mlrPoIFN-α可分别抑制PRV和PPV对PK-15细胞株致细胞病变效应。[结论]rPoIFN-α具有IFN-α的基本理化性状,其在体外可分别抑制PRV、PPV对PK-15细胞株致细胞病变效应,但剂量有差别。[Objective] The research aimed to test and identify the physicochemical characters and antiviral activity in vitro of semi-finished product of the recombinant porcine rPoIFN-α.[Method] HEp-2/VSV system was used to test the antiviral activity of three batches of rPoIFN-α protein.Using recombinant human IFN-α as reference,the titer of interferon was measured.The semi-finished product of rPoIFN-α with the known titer were treated with 0.25% trypsin,HCl and mouse anti-porcine IFN-α monoclonal antibody.And the anti-viral activity of each batch of rPoIFN-α was detected.The physicochemical characters of rPoIFN-α were evaluated.The inhibition of induced cytopathic effect of rPoIFN-αon PPV （Porcine parvovirus） and PRV （Porcine pseudorabies） on swine kidney cell （PK-15） was determined.And the antiviral activity of rPoIFN-α in vitro was observed.[Result] The titers of semi-finished products of rPoIFN-α titrated by HEp-2/VSV system could reach 1.5×105 IU/ml,with the specific activity of 1.1×106 IU/mg.The residual rate of the tier of rPoIFN-α treated by 0.25% trypsin at 37 ℃ for 1 h was less than 1%.And that treated with HCl （pH=2.0） for 72 h was up to 95%.And that treated at 56 ℃ for 30 min and that treated by mouse anti-porcine IFN-α monoclonal antibody at 37 ℃for 1 h were higher than 47% and about 1% respectively.The antiviral test in vitro showed that 50 and 500 IU/ml rPoIFN-α could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines.[Conclusion] rPoIFN-α had the basic physicochemical characters of IFN-α.And it could inhibit the induced cytopathic effect of PRV and PPV on PK-15 cell lines,but there was dosage difference.

关 键 词：猪Α干扰素 理化性状 抗病毒活性 

分 类 号：S858.28[农业科学—临床兽医学]