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 |
| Published: |
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 |
| Summary: | 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 |
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| ISSN: | 2539-6056 2539-6056 |