CAJ | 학술논문

[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1.5×10~5 IU/ml,比活性达1.1×10~6 IU/mg;rPoIFN-α经0.25%胰蛋白酶37 ℃作用1 h,效价残留率低于1%,经HCl(pH=2.0)处理72 h效价残留率高达95%以上,经56 ℃处理30 min效价残留率高于47%,经鼠抗猪α干扰素单克隆抗体37 ℃作用1 h后效价残留率约为1%;体外抗病毒试验表明,用50和500 IU/ml rPoIFN-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.
[목적]연구중조저α간우소(rPoIFN-α)반성품이화성상병대기체외항병독활성진행측시여감정.[방법]용HEp-2/VSV체계대3비rPoIFN-α단백진행항병독활성검측,이중조인IFN-α위삼비품,측정간우소효개;용0.25%이단백매HCl이급서항저α간우소단극륭항체작용이지효개적rPoIFN-α반성품,병검측각비차항병독활성,대rPoIFN-α이화성상진행평개;재저신세포주(PK-15)상검측rPoIFN-α대저세소병독(PPV)화저위광견병독(PRV)적치세포병변억제효응,관찰rPoIFN-α적체외항병독활성.[결과]HEp-2/VSV체계적정rPoIFN-α반성품효개가체1.5×10~5 IU/ml,비활성체1.1×10~6 IU/mg;rPoIFN-α경0.25%이단백매37 ℃작용1 h,효개잔류솔저우1%,경HCl(pH=2.0)처리72 h효개잔류솔고체95%이상,경56 ℃처리30 min효개잔류솔고우47%,경서항저α간우소단극륭항체37 ℃작용1 h후효개잔류솔약위1%;체외항병독시험표명,용50화500 IU/ml rPoIFN-α가분별억제PRV화PPV대PK-15세포주치세포병변효응.[결론]rPoIFN-α구유IFN-α적기본이화성상,기재체외가분별억제PRV、PPV대PK-15세포주치세포병변효응,단제량유차별.
[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 1.5×10~5 IU/ml, with the specific activity of 1.1×10~6 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.