Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography
Nucleocapsid (N) protein of Nipah virus (NiV) is a potential serological marker used in the diagnosis of NiV infections. In this study, a rapid and efficient purification system, HisTrapTM 6 Fast Flowpacked bed column was applied to purify recombinant histidine-tagged N protein of NiV from clarifi...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English English |
| Published: |
Elsevier Ltd.
2009
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| Online Access: | http://psasir.upm.edu.my/id/eprint/16395/ http://psasir.upm.edu.my/id/eprint/16395/1/Purification%20of%20histidine.pdf |
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| author | Fui , Chin Chong Wen, Siang Tan Awang Biak, Dayang Radiah Tau, Chuan Ling Beng , Ti Tey |
| author_facet | Fui , Chin Chong Wen, Siang Tan Awang Biak, Dayang Radiah Tau, Chuan Ling Beng , Ti Tey |
| author_sort | Fui , Chin Chong |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Nucleocapsid (N) protein of Nipah virus (NiV) is a potential serological marker used in the diagnosis of NiV
infections. In this study, a rapid and efficient purification system, HisTrapTM 6 Fast Flowpacked bed column
was applied to purify recombinant histidine-tagged N protein of NiV from clarified feedstock. The optimizations
of binding and elution conditions of N protein of NiV onto and from Nickel SepharoseTM 6 Fast Flow were investigated. The optimal binding was achieved at pH 7.5, superficial velocity of 1.25 cm/min.The bound N protein was successfully recovered by a stepwise elution with different concentration of imidazole (50, 150, 300 and 500 mM). The N protein of NiV was captured and eluted from an inlet N protein concentration of 0.4 mg/ml in a scale-up immobilizedmetal affinity chromatography (IMAC) packed
bed column of Nickel SepharoseTM 6 Fast Flow with the optimized condition obtained from the method scouting. The purification of histidine-tagged N protein using IMAC packed bed column has resulted a 68.3% yield and a purification factor of 7.94. |
| first_indexed | 2025-11-15T08:07:16Z |
| format | Article |
| id | upm-16395 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:07:16Z |
| publishDate | 2009 |
| publisher | Elsevier Ltd. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-163952015-10-05T07:28:10Z http://psasir.upm.edu.my/id/eprint/16395/ Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography Fui , Chin Chong Wen, Siang Tan Awang Biak, Dayang Radiah Tau, Chuan Ling Beng , Ti Tey Nucleocapsid (N) protein of Nipah virus (NiV) is a potential serological marker used in the diagnosis of NiV infections. In this study, a rapid and efficient purification system, HisTrapTM 6 Fast Flowpacked bed column was applied to purify recombinant histidine-tagged N protein of NiV from clarified feedstock. The optimizations of binding and elution conditions of N protein of NiV onto and from Nickel SepharoseTM 6 Fast Flow were investigated. The optimal binding was achieved at pH 7.5, superficial velocity of 1.25 cm/min.The bound N protein was successfully recovered by a stepwise elution with different concentration of imidazole (50, 150, 300 and 500 mM). The N protein of NiV was captured and eluted from an inlet N protein concentration of 0.4 mg/ml in a scale-up immobilizedmetal affinity chromatography (IMAC) packed bed column of Nickel SepharoseTM 6 Fast Flow with the optimized condition obtained from the method scouting. The purification of histidine-tagged N protein using IMAC packed bed column has resulted a 68.3% yield and a purification factor of 7.94. Elsevier Ltd. 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16395/1/Purification%20of%20histidine.pdf Fui , Chin Chong and Wen, Siang Tan and Awang Biak, Dayang Radiah and Tau, Chuan Ling and Beng , Ti Tey (2009) Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 877 (14-15). pp. 1561-1567. ISSN 1570- 0232 Plant viruses Proteins - Purification Chromatographic analysis doi:10.1016/j.jchromb.2009.03.048 English |
| spellingShingle | Plant viruses Proteins - Purification Chromatographic analysis Fui , Chin Chong Wen, Siang Tan Awang Biak, Dayang Radiah Tau, Chuan Ling Beng , Ti Tey Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title | Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title_full | Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title_fullStr | Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title_full_unstemmed | Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title_short | Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography |
| title_sort | purification of histidine-tagged nucleocapsid protein of nipah virus using immobilized metal affinity chromatography |
| topic | Plant viruses Proteins - Purification Chromatographic analysis |
| url | http://psasir.upm.edu.my/id/eprint/16395/ http://psasir.upm.edu.my/id/eprint/16395/ http://psasir.upm.edu.my/id/eprint/16395/1/Purification%20of%20histidine.pdf |