[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 tes...[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. 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-α on 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 of 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.展开更多
基金Supported by Natural Science Research Project of Anhui Educational Committee (2004kj218 )Major Special Program of Science and Technology Grand Plan of Anhui Province (08010302179)~~
文摘[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. 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-α on 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 of 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.
