Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research

BMC Microbiology

Research Article

Lan Zhang¹  Eberhard Durr¹  James C. Cook¹  Craig T. Przysiecki¹  Zhiyun Wen¹  Melissa A. Boddicker¹  Debbie D. Nahas¹  Amy S. Espeseth¹  Robin M. Kaufhold¹  Sinoeun Touch¹  Puneet Khandelwal¹  Julie M. Skinner¹  Bob J. Lucas¹  Ping Qiu² 

[1]Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA

[2]Translational Molecular Biomarkers (Rahway, NJ), MRL, Merck & Co., Inc, Kenilworth, NJ, USA

关键词: Chlamydia trachomatis;    MOMP;    Outer membrane expression;    Immunogenicity;   

DOI  :  10.1186/s12866-016-0787-3

 received in 2016-03-10, accepted in 2016-07-22,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundChlamydia 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.ResultsWe 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).ConclusionsC. 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.

【 授权许可】

CC BY   
© The Author(s). 2016