The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein
The quantitative determination of recombinant protein in crude protein is not commonly used because the protein is not in purified form. The Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was developed for quantitative determination of protein. This method has been use for prot...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Chaing Mai University
2013-05-01
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| Series: | Journal of Associated Medical Sciences |
| Subjects: | |
| Online Access: | https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/60006 |
| _version_ | 1811199630365425664 |
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| author | Kannaporn Intachai Chanyanuch Putim Watchara Kasinrerk Chatchai Tayapiwatana Bordin Butr-Indr |
| author_facet | Kannaporn Intachai Chanyanuch Putim Watchara Kasinrerk Chatchai Tayapiwatana Bordin Butr-Indr |
| author_sort | Kannaporn Intachai |
| collection | DOAJ |
| description | The quantitative determination of recombinant protein in crude protein is not commonly used because the protein is not in purified form. The Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was developed for quantitative determination of protein. This method has been use for protein quantitative determinations in many propose such as Tamm-Horsfall glycoprotein determination in kidney stone disease patient and determination of titin and nebulin in chicken meat product for meat industries. In this study, the method of quantitative determination of extracellular recombinant single-chain variable fragment anti-HIV-1 p17 proteins (scFv anti-p17) from Escherichia coli stain HB2151 by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was developed by the application of bovine serum albumin as external and internal standard. The range of detection was 1.875 - 0.234 μg. These results presented variation coefficient less than 35%. The method applied to quantitative determination of scFv anti-p17 in 19 of crude protein samples. The quantitative protein concentration results related to the enzyme-linked immunosorbent assay (ELISA) and western immunoblotting method. The quantitative SDS-PAGE method is capable for quantitative determination of known protein from crude protein extraction. This method decreased the purification step, time consuming and protein yield loss. This application is capable to apply with the other pathogenic protein markers in the future clinical applications. Bull Chiang Mai Assoc Med Sci 2013; 46(2): 107-121 |
| first_indexed | 2024-04-12T01:50:36Z |
| format | Article |
| id | doaj.art-8ceb5cf0f3a544e2bf6ea7c45f2bd5da |
| institution | Directory Open Access Journal |
| issn | 2539-6056 2539-6056 |
| language | English |
| last_indexed | 2024-04-12T01:50:36Z |
| publishDate | 2013-05-01 |
| publisher | Chaing Mai University |
| record_format | Article |
| series | Journal of Associated Medical Sciences |
| spelling | doaj.art-8ceb5cf0f3a544e2bf6ea7c45f2bd5da2022-12-22T03:52:56ZengChaing Mai UniversityJournal of Associated Medical Sciences2539-60562539-60562013-05-0146210710760006The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude proteinKannaporn Intachai0Chanyanuch Putim1Watchara Kasinrerk2Chatchai Tayapiwatana3Bordin Butr-Indr41 Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 2 Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 3 Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University1 Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 2 Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 3 Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Develop¬ment Agency at the Faculty of Associated Medical Sciences, Chiang Mai University1 Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 2 Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 3 Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Develop¬ment Agency at the Faculty of Associated Medical Sciences, Chiang Mai University1 Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 2 Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 3 Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Develop¬ment Agency at the Faculty of Associated Medical Sciences, Chiang Mai University1 Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 2 Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Science, Chiang Mai University 3 Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Develop¬ment Agency at the Faculty of Associated Medical Sciences, Chiang Mai UniversityThe quantitative determination of recombinant protein in crude protein is not commonly used because the protein is not in purified form. The Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was developed for quantitative determination of protein. This method has been use for protein quantitative determinations in many propose such as Tamm-Horsfall glycoprotein determination in kidney stone disease patient and determination of titin and nebulin in chicken meat product for meat industries. In this study, the method of quantitative determination of extracellular recombinant single-chain variable fragment anti-HIV-1 p17 proteins (scFv anti-p17) from Escherichia coli stain HB2151 by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was developed by the application of bovine serum albumin as external and internal standard. The range of detection was 1.875 - 0.234 μg. These results presented variation coefficient less than 35%. The method applied to quantitative determination of scFv anti-p17 in 19 of crude protein samples. The quantitative protein concentration results related to the enzyme-linked immunosorbent assay (ELISA) and western immunoblotting method. The quantitative SDS-PAGE method is capable for quantitative determination of known protein from crude protein extraction. This method decreased the purification step, time consuming and protein yield loss. This application is capable to apply with the other pathogenic protein markers in the future clinical applications. Bull Chiang Mai Assoc Med Sci 2013; 46(2): 107-121https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/60006SDS-PAGEscFv anti-p17HIV-1 |
| spellingShingle | Kannaporn Intachai Chanyanuch Putim Watchara Kasinrerk Chatchai Tayapiwatana Bordin Butr-Indr The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein Journal of Associated Medical Sciences SDS-PAGE scFv anti-p17 HIV-1 |
| title | The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein |
| title_full | The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein |
| title_fullStr | The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein |
| title_full_unstemmed | The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein |
| title_short | The application of quantitative SDS-PAGE method for measurement of extracellular recombinant scFv anti-p17 in crude protein |
| title_sort | application of quantitative sds page method for measurement of extracellular recombinant scfv anti p17 in crude protein |
| topic | SDS-PAGE scFv anti-p17 HIV-1 |
| url | https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/60006 |
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