Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells
Intracellular green fluorescent protein (GFP) can be separated and purified from intact Escherichia coli cells by a preparative native polyacrylamide gel electrophoresis (n-PAGE). The effects of operating parameters such as the volume and concentration of feedstock and the pore size and height of re...
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Elsevier
2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/23105/ http://psasir.upm.edu.my/id/eprint/23105/1/Optimization%20of%20a%20native%20gel%20electrophoretic%20process%20for%20the%20purification%20of%20intracellular%20green%20fluorescent%20protein%20from%20intact%20Escherichia%20coli%20cells.pdf |
| _version_ | 1848844664488067072 |
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| author | Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti |
| author_facet | Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti |
| author_sort | Chew, Few Ne |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Intracellular green fluorescent protein (GFP) can be separated and purified from intact Escherichia coli cells by a preparative native polyacrylamide gel electrophoresis (n-PAGE). The effects of operating parameters such as the volume and concentration of feedstock and the pore size and height of resolving gel on the purity and yield of GFP were studied using a 1.7 cm internal diameter gel column. The optimum conditions for this preparative n-PAGE operation were determined to be 100 μl of 15% (w/v) feedstock and a 12% (w/v) resolving gel with a gel height of 2 cm. The purity and yield of the recovered GFP were 98 and 88%, respectively. The results of scalability studies show that the ratio of feedstock volume to cross-sectional area of the column is an important consideration for scaling up the preparative n-PAGE. |
| first_indexed | 2025-11-15T08:34:31Z |
| format | Article |
| id | upm-23105 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T08:34:31Z |
| publishDate | 2011 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-231052015-12-07T02:47:21Z http://psasir.upm.edu.my/id/eprint/23105/ Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti Intracellular green fluorescent protein (GFP) can be separated and purified from intact Escherichia coli cells by a preparative native polyacrylamide gel electrophoresis (n-PAGE). The effects of operating parameters such as the volume and concentration of feedstock and the pore size and height of resolving gel on the purity and yield of GFP were studied using a 1.7 cm internal diameter gel column. The optimum conditions for this preparative n-PAGE operation were determined to be 100 μl of 15% (w/v) feedstock and a 12% (w/v) resolving gel with a gel height of 2 cm. The purity and yield of the recovered GFP were 98 and 88%, respectively. The results of scalability studies show that the ratio of feedstock volume to cross-sectional area of the column is an important consideration for scaling up the preparative n-PAGE. Elsevier 2011-01 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/23105/1/Optimization%20of%20a%20native%20gel%20electrophoretic%20process%20for%20the%20purification%20of%20intracellular%20green%20fluorescent%20protein%20from%20intact%20Escherichia%20coli%20cells.pdf Chew, Few Ne and Tan, Wen Siang and Ling, Tau Chuan and Tey, Beng Ti (2011) Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells. Process Biochemistry, 46 (1). pp. 399-403. ISSN 1359-5113; ESSN: 1873-3298 10.1016/j.procbio.2010.07.032 |
| spellingShingle | Chew, Few Ne Tan, Wen Siang Ling, Tau Chuan Tey, Beng Ti Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title | Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title_full | Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title_fullStr | Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title_full_unstemmed | Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title_short | Optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact Escherichia coli cells |
| title_sort | optimization of a native gel electrophoretic process for the purification of intracellular green fluorescent protein from intact escherichia coli cells |
| url | http://psasir.upm.edu.my/id/eprint/23105/ http://psasir.upm.edu.my/id/eprint/23105/ http://psasir.upm.edu.my/id/eprint/23105/1/Optimization%20of%20a%20native%20gel%20electrophoretic%20process%20for%20the%20purification%20of%20intracellular%20green%20fluorescent%20protein%20from%20intact%20Escherichia%20coli%20cells.pdf |