Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity
Specific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most...
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| Format: | Article |
| Language: | Russian |
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Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products»
2020-03-01
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| Series: | Биопрепараты: Профилактика, диагностика, лечение |
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| Online Access: | https://www.biopreparations.ru/jour/article/view/249 |
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| author | M. L. Baykova I. M. Shcherbachenko L. A. Gayderova O. B. Ustinnikova A. A. Movsesyants |
| author_facet | M. L. Baykova I. M. Shcherbachenko L. A. Gayderova O. B. Ustinnikova A. A. Movsesyants |
| author_sort | M. L. Baykova |
| collection | DOAJ |
| description | Specific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most appropriate conditions for in vitro determination of IFN-α product specific activity. Materials and methods: Vero, MDBK, Hep-2, and A-549 homologous and heterologous cell cultures, as well as vesicular stomatitis Indiana virus (VSV) and murine encephalomyocarditis (EMC) virus at a dose of 100 TCD50 /0.1 mL were used for determination of specific antiviral activity. The international reference standard of recombinant interferon alpha-2b activity (Interferon alpha 2b, human, rDNA, E. coli-derived, 2nd WHO International Standard, 1999, NIBSC Code No. 95/566) and human recombinant interferon alpha 2b in the form of solution (batch No. 040214, Pharmapark LLC, Russia) were used as IFN-α samples. Results: the analysis of the obtained data helped to determine: the combinations of cell lines and the indicator virus most sensitive to IFN-α; the optimal concentration of fetal serum in the medium, and the optimal time parameters; the preferred method of reporting test results. Conclusions: the following test conditions were found to be optimal: the MDBK/VSV and Hep-2/EMC combinations proved to be the most sensitive to IFN-α; the optimal period of interferon and cell culture incubation—24 hours; the optimal concentration of fetal bovine serum in the culture medium used for diluting interferon products—2–5%. The instrumental procedure is preferred for reporting the results of interferon antiviral activity determination, because it is up-to-date, reliable, accurate and time-efficient. |
| format | Article |
| id | doaj-art-a35ef6aa98554ff8a13b27413037261d |
| institution | Kabale University |
| issn | 2221-996X 2619-1156 |
| language | Russian |
| publishDate | 2020-03-01 |
| publisher | Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products» |
| record_format | Article |
| series | Биопрепараты: Профилактика, диагностика, лечение |
| spelling | doaj-art-a35ef6aa98554ff8a13b27413037261d2025-08-20T03:59:39ZrusMinistry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products»Биопрепараты: Профилактика, диагностика, лечение2221-996X2619-11562020-03-01201687310.30895/2221-996X-2020-20-1-68-73190Aspects of a Methodological Approach to Determination of Interferon Alpha Specific ActivityM. L. Baykova0I. M. Shcherbachenko1L. A. Gayderova2O. B. Ustinnikova3A. A. Movsesyants4Scientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsScientific Centre for Expert Evaluation of Medicinal ProductsSpecific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most appropriate conditions for in vitro determination of IFN-α product specific activity. Materials and methods: Vero, MDBK, Hep-2, and A-549 homologous and heterologous cell cultures, as well as vesicular stomatitis Indiana virus (VSV) and murine encephalomyocarditis (EMC) virus at a dose of 100 TCD50 /0.1 mL were used for determination of specific antiviral activity. The international reference standard of recombinant interferon alpha-2b activity (Interferon alpha 2b, human, rDNA, E. coli-derived, 2nd WHO International Standard, 1999, NIBSC Code No. 95/566) and human recombinant interferon alpha 2b in the form of solution (batch No. 040214, Pharmapark LLC, Russia) were used as IFN-α samples. Results: the analysis of the obtained data helped to determine: the combinations of cell lines and the indicator virus most sensitive to IFN-α; the optimal concentration of fetal serum in the medium, and the optimal time parameters; the preferred method of reporting test results. Conclusions: the following test conditions were found to be optimal: the MDBK/VSV and Hep-2/EMC combinations proved to be the most sensitive to IFN-α; the optimal period of interferon and cell culture incubation—24 hours; the optimal concentration of fetal bovine serum in the culture medium used for diluting interferon products—2–5%. The instrumental procedure is preferred for reporting the results of interferon antiviral activity determination, because it is up-to-date, reliable, accurate and time-efficient.https://www.biopreparations.ru/jour/article/view/249interferon alpha (ifn-α)quality controlbioassayspecific antiviral activity |
| spellingShingle | M. L. Baykova I. M. Shcherbachenko L. A. Gayderova O. B. Ustinnikova A. A. Movsesyants Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity Биопрепараты: Профилактика, диагностика, лечение interferon alpha (ifn-α) quality control bioassay specific antiviral activity |
| title | Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity |
| title_full | Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity |
| title_fullStr | Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity |
| title_full_unstemmed | Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity |
| title_short | Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity |
| title_sort | aspects of a methodological approach to determination of interferon alpha specific activity |
| topic | interferon alpha (ifn-α) quality control bioassay specific antiviral activity |
| url | https://www.biopreparations.ru/jour/article/view/249 |
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