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High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli
BACKGROUND: Helicobacter pylori neutrophil-activating protein (HP-NAP) is involved in H. pylori-induced gastric inflammation. Due to its immunogenic and immunomodulatory properties, HP-NAP has been used for developing vaccines against H. pylori infection and new drugs for cancer therapy. RESULTS: He...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4425898/ https://www.ncbi.nlm.nih.gov/pubmed/25880121 http://dx.doi.org/10.1186/s12896-015-0136-x |
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author | Yang, Yu-Chi Kuo, Ting-Yu Hong, Zhi-Wei Chang, Han-Wen Chen, Chung-Chu Tsai, Te-Lung Fu, Hua-Wen |
author_facet | Yang, Yu-Chi Kuo, Ting-Yu Hong, Zhi-Wei Chang, Han-Wen Chen, Chung-Chu Tsai, Te-Lung Fu, Hua-Wen |
author_sort | Yang, Yu-Chi |
collection | PubMed |
description | BACKGROUND: Helicobacter pylori neutrophil-activating protein (HP-NAP) is involved in H. pylori-induced gastric inflammation. Due to its immunogenic and immunomodulatory properties, HP-NAP has been used for developing vaccines against H. pylori infection and new drugs for cancer therapy. RESULTS: Here, we provide a simple process for high-yield production of HP-NAP by applying one-step negative chromatography to purify recombinant HP-NAP expressed in Escherichia coli (E. coli). In our E. coli expression system, recombinant HP-NAP constitutes nearly 70% of the total protein. Overexpressed recombinant HP-NAP is almost completely soluble upon cell lysis at pH 9.5. Under the optimal condition at pH 8.0, recombinant HP-NAP with purity higher than 95% can be obtained from E. coli by collecting the unbound fraction using diethylaminoethyl (DEAE) Sephadex resin in batch mode. The overall yield of HP-NAP from a 50-ml E. coli culture is ~19 mg. The purified HP-NAP folds into a multimer with a secondary structure of α-helix and is able to trigger the production of reactive oxygen species by neutrophils. CONCLUSIONS: Purification of recombinant HP-NAP overexpressed in E. coli using DEAE Sephadex negative mode batch chromatography is an efficient method for high-yield production of highly pure HP-NAP in its native state. The purified HP-NAP is useful for various clinical applications including vaccine development, diagnosis, and new drug development. |
format | Online Article Text |
id | pubmed-4425898 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44258982015-05-10 High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli Yang, Yu-Chi Kuo, Ting-Yu Hong, Zhi-Wei Chang, Han-Wen Chen, Chung-Chu Tsai, Te-Lung Fu, Hua-Wen BMC Biotechnol Research Article BACKGROUND: Helicobacter pylori neutrophil-activating protein (HP-NAP) is involved in H. pylori-induced gastric inflammation. Due to its immunogenic and immunomodulatory properties, HP-NAP has been used for developing vaccines against H. pylori infection and new drugs for cancer therapy. RESULTS: Here, we provide a simple process for high-yield production of HP-NAP by applying one-step negative chromatography to purify recombinant HP-NAP expressed in Escherichia coli (E. coli). In our E. coli expression system, recombinant HP-NAP constitutes nearly 70% of the total protein. Overexpressed recombinant HP-NAP is almost completely soluble upon cell lysis at pH 9.5. Under the optimal condition at pH 8.0, recombinant HP-NAP with purity higher than 95% can be obtained from E. coli by collecting the unbound fraction using diethylaminoethyl (DEAE) Sephadex resin in batch mode. The overall yield of HP-NAP from a 50-ml E. coli culture is ~19 mg. The purified HP-NAP folds into a multimer with a secondary structure of α-helix and is able to trigger the production of reactive oxygen species by neutrophils. CONCLUSIONS: Purification of recombinant HP-NAP overexpressed in E. coli using DEAE Sephadex negative mode batch chromatography is an efficient method for high-yield production of highly pure HP-NAP in its native state. The purified HP-NAP is useful for various clinical applications including vaccine development, diagnosis, and new drug development. BioMed Central 2015-04-08 /pmc/articles/PMC4425898/ /pubmed/25880121 http://dx.doi.org/10.1186/s12896-015-0136-x Text en © Yang et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Yang, Yu-Chi Kuo, Ting-Yu Hong, Zhi-Wei Chang, Han-Wen Chen, Chung-Chu Tsai, Te-Lung Fu, Hua-Wen High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title | High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title_full | High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title_fullStr | High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title_full_unstemmed | High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title_short | High yield purification of Helicobacter pylori neutrophil-activating protein overexpressed in Escherichia coli |
title_sort | high yield purification of helicobacter pylori neutrophil-activating protein overexpressed in escherichia coli |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4425898/ https://www.ncbi.nlm.nih.gov/pubmed/25880121 http://dx.doi.org/10.1186/s12896-015-0136-x |
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