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Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research
BACKGROUND: Chlamydia trachomatis is a human pathogen which causes a number of pathologies, including genital tract infections in women that can result in tubal infertility. Prevention of infection and disease control might be achieved through vaccination; however, a safe, efficacious and cost-effec...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4963994/ https://www.ncbi.nlm.nih.gov/pubmed/27464881 http://dx.doi.org/10.1186/s12866-016-0787-3 |
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author | Wen, Zhiyun Boddicker, Melissa A. Kaufhold, Robin M. Khandelwal, Puneet Durr, Eberhard Qiu, Ping Lucas, Bob J. Nahas, Debbie D. Cook, James C. Touch, Sinoeun Skinner, Julie M. Espeseth, Amy S. Przysiecki, Craig T. Zhang, Lan |
author_facet | Wen, Zhiyun Boddicker, Melissa A. Kaufhold, Robin M. Khandelwal, Puneet Durr, Eberhard Qiu, Ping Lucas, Bob J. Nahas, Debbie D. Cook, James C. Touch, Sinoeun Skinner, Julie M. Espeseth, Amy S. Przysiecki, Craig T. Zhang, Lan |
author_sort | Wen, Zhiyun |
collection | PubMed |
description | BACKGROUND: Chlamydia trachomatis is a human pathogen which causes a number of pathologies, including genital tract infections in women that can result in tubal infertility. Prevention of infection and disease control might be achieved through vaccination; however, a safe, efficacious and cost-effective vaccine against C. trachomatis infection remains an unmet medical need. C. trachomatis major outer membrane protein (MOMP), a β-barrel integral outer membrane protein, is the most abundant antigen in the outer membrane of the bacterium and has been evaluated as a subunit vaccine candidate. Recombinant MOMP (rMOMP) expressed in E. coli cytoplasm forms inclusion bodies and rMOMP extracted from inclusion bodies results in a reduced level of protection compared to the native MOMP in a mouse challenge model. RESULTS: We sought to target the recombinant expression of MOMP to the E. coli outer membrane (OM). Successful surface expression was achieved with codon harmonization, utilization of low copy number vectors and promoters with moderate strength, suitable leader sequences and optimization of cell culture conditions. rMOMP was extracted from E. coli outer membrane, purified, and characterized biophysically. The OM expressed and purified rMOMP is immunogenic in mice and elicits antibodies that react to the native antigen, Chlamydia elementary body (EB). CONCLUSIONS: C. trachomatis MOMP was functionally expressed on the surface of E. coli outer membrane. The OM expressed and purified rMOMP elicits antibodies that react to the native antigen, Chlamydia EB, in a mouse immunogenicity model. Surface expression of MOMP could provide useful reagents for vaccine research, and the methodology could serve as a platform to produce other outer membrane proteins recombinantly. |
format | Online Article Text |
id | pubmed-4963994 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-49639942016-07-29 Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research Wen, Zhiyun Boddicker, Melissa A. Kaufhold, Robin M. Khandelwal, Puneet Durr, Eberhard Qiu, Ping Lucas, Bob J. Nahas, Debbie D. Cook, James C. Touch, Sinoeun Skinner, Julie M. Espeseth, Amy S. Przysiecki, Craig T. Zhang, Lan BMC Microbiol Research Article BACKGROUND: Chlamydia trachomatis is a human pathogen which causes a number of pathologies, including genital tract infections in women that can result in tubal infertility. Prevention of infection and disease control might be achieved through vaccination; however, a safe, efficacious and cost-effective vaccine against C. trachomatis infection remains an unmet medical need. C. trachomatis major outer membrane protein (MOMP), a β-barrel integral outer membrane protein, is the most abundant antigen in the outer membrane of the bacterium and has been evaluated as a subunit vaccine candidate. Recombinant MOMP (rMOMP) expressed in E. coli cytoplasm forms inclusion bodies and rMOMP extracted from inclusion bodies results in a reduced level of protection compared to the native MOMP in a mouse challenge model. RESULTS: We sought to target the recombinant expression of MOMP to the E. coli outer membrane (OM). Successful surface expression was achieved with codon harmonization, utilization of low copy number vectors and promoters with moderate strength, suitable leader sequences and optimization of cell culture conditions. rMOMP was extracted from E. coli outer membrane, purified, and characterized biophysically. The OM expressed and purified rMOMP is immunogenic in mice and elicits antibodies that react to the native antigen, Chlamydia elementary body (EB). CONCLUSIONS: C. trachomatis MOMP was functionally expressed on the surface of E. coli outer membrane. The OM expressed and purified rMOMP elicits antibodies that react to the native antigen, Chlamydia EB, in a mouse immunogenicity model. Surface expression of MOMP could provide useful reagents for vaccine research, and the methodology could serve as a platform to produce other outer membrane proteins recombinantly. BioMed Central 2016-07-27 /pmc/articles/PMC4963994/ /pubmed/27464881 http://dx.doi.org/10.1186/s12866-016-0787-3 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Wen, Zhiyun Boddicker, Melissa A. Kaufhold, Robin M. Khandelwal, Puneet Durr, Eberhard Qiu, Ping Lucas, Bob J. Nahas, Debbie D. Cook, James C. Touch, Sinoeun Skinner, Julie M. Espeseth, Amy S. Przysiecki, Craig T. Zhang, Lan Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title | Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title_full | Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title_fullStr | Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title_full_unstemmed | Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title_short | Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research |
title_sort | recombinant expression of chlamydia trachomatis major outer membrane protein in e. coli outer membrane as a substrate for vaccine research |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4963994/ https://www.ncbi.nlm.nih.gov/pubmed/27464881 http://dx.doi.org/10.1186/s12866-016-0787-3 |
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