IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL
The article is devoted to develop of an approach for the identification of new stimulator of ematopoiesis, EPO-Fc fusion protein, which is banned by the World Anti-doping Agency (WADA) to use by athletes since it has become doping. Existing methods of qualitative determination of this substances in...
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| Format: | Article |
| Language: | Russian |
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MIREA - Russian Technological University
2016-10-01
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| Series: | Тонкие химические технологии |
| Subjects: | |
| Online Access: | https://www.finechem-mirea.ru/jour/article/view/51 |
| _version_ | 1826565738792484864 |
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| author | P. V. Postnikov G. I. Krotov Yu. A. Efimova M. A. Dikunets |
| author_facet | P. V. Postnikov G. I. Krotov Yu. A. Efimova M. A. Dikunets |
| author_sort | P. V. Postnikov |
| collection | DOAJ |
| description | The article is devoted to develop of an approach for the identification of new stimulator of ematopoiesis, EPO-Fc fusion protein, which is banned by the World Anti-doping Agency (WADA) to use by athletes since it has become doping. Existing methods of qualitative determination of this substances in routine practice of antidoping laboratories such as polyacrylamide gelelectrophoresis in presence of sodium dodecylsulphate (SDS-PAGE) or lauroylsarcosinate (SARPAGE) are insufficiently specific. The article shows the principal possibility of identification of EPO-Fc fusion protein by means of IEF-PAGE in carrier ampholyte-based gels with a pH range 2-6 after Fc-fragment removal via fermentative hydrolysis.It has been shown that the removing of the crystallizable fragment leads to decrease of molecular weight of whole hybrid molecule and to increase its electrophoretic mobility that allows to detect this banned substances with high specificity by existing methods. During the study the enzyme for hydrolytic cleavage and optimum conditions of hydrolysis of EPO-Fc in serum samples were selected. |
| first_indexed | 2024-04-10T03:28:59Z |
| format | Article |
| id | doaj.art-b8f9eeffdaaf47e9a34dfe7308eb1c82 |
| institution | Directory Open Access Journal |
| issn | 2410-6593 2686-7575 |
| language | Russian |
| last_indexed | 2025-03-14T10:40:32Z |
| publishDate | 2016-10-01 |
| publisher | MIREA - Russian Technological University |
| record_format | Article |
| series | Тонкие химические технологии |
| spelling | doaj.art-b8f9eeffdaaf47e9a34dfe7308eb1c822025-03-02T10:57:34ZrusMIREA - Russian Technological UniversityТонкие химические технологии2410-65932686-75752016-10-01115708010.32362/2410-6593-2016-11-5-70-8051IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROLP. V. Postnikov0G. I. Krotov1Yu. A. Efimova2M. A. Dikunets3Federal State Budgetary Institution “Antidoping Centre”Federal State Budgetary Institution “Antidoping Centre”Moscow Technological University (Institute of Fine Chemical Technologies)Federal State Budgetary Institution “Antidoping Centre”The article is devoted to develop of an approach for the identification of new stimulator of ematopoiesis, EPO-Fc fusion protein, which is banned by the World Anti-doping Agency (WADA) to use by athletes since it has become doping. Existing methods of qualitative determination of this substances in routine practice of antidoping laboratories such as polyacrylamide gelelectrophoresis in presence of sodium dodecylsulphate (SDS-PAGE) or lauroylsarcosinate (SARPAGE) are insufficiently specific. The article shows the principal possibility of identification of EPO-Fc fusion protein by means of IEF-PAGE in carrier ampholyte-based gels with a pH range 2-6 after Fc-fragment removal via fermentative hydrolysis.It has been shown that the removing of the crystallizable fragment leads to decrease of molecular weight of whole hybrid molecule and to increase its electrophoretic mobility that allows to detect this banned substances with high specificity by existing methods. During the study the enzyme for hydrolytic cleavage and optimum conditions of hydrolysis of EPO-Fc in serum samples were selected.https://www.finechem-mirea.ru/jour/article/view/51fermentative hydrolysisepo-fc fusion proteinelectrophoresisepo-hinge fragmentantidoping control |
| spellingShingle | P. V. Postnikov G. I. Krotov Yu. A. Efimova M. A. Dikunets IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL Тонкие химические технологии fermentative hydrolysis epo-fc fusion protein electrophoresis epo-hinge fragment antidoping control |
| title | IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL |
| title_full | IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL |
| title_fullStr | IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL |
| title_full_unstemmed | IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL |
| title_short | IDENTIFICATION OF EPO-Fс FUSION PROTEIN BY MEANS OF POLYACRYLAMIDE GEL-ELECTROPHORESIS WITH ISOELECTROFOCUSING (IEF-PAGE)AND IN PRESENCE OF SODIUM DODECYLSULPHATE (SDS-PAGE)/ LAUROYLSARCOSINATE (SAR-PAGE) FOR THE PURPOSE OF ANTI-DOPING CONTROL |
| title_sort | identification of epo fс fusion protein by means of polyacrylamide gel electrophoresis with isoelectrofocusing ief page and in presence of sodium dodecylsulphate sds page lauroylsarcosinate sar page for the purpose of anti doping control |
| topic | fermentative hydrolysis epo-fc fusion protein electrophoresis epo-hinge fragment antidoping control |
| url | https://www.finechem-mirea.ru/jour/article/view/51 |
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