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Recombinant porcine interferon-gamma expressed in CHO cells and its antiviral activity / 生物工程学报
Chinese Journal of Biotechnology ; (12): 4784-4795, 2023.
Article in Zh | WPRIM | ID: wpr-1008058
Responsible library: WPRO
ABSTRACT
The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×107 U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.
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Full text: 1 Database: WPRIM Main subject: Antiviral Agents / Sincalide / Swine / Recombinant Proteins / Cricetulus / Interferon-gamma / CHO Cells Limits: Animals Language: Zh Journal: Chinese Journal of Biotechnology Year: 2023 Document type: Article
Full text: 1 Database: WPRIM Main subject: Antiviral Agents / Sincalide / Swine / Recombinant Proteins / Cricetulus / Interferon-gamma / CHO Cells Limits: Animals Language: Zh Journal: Chinese Journal of Biotechnology Year: 2023 Document type: Article