title: Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research.
abstract: 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.
  
  
  
  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).
  
  
  
  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.
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Zhiyun
authors.initials: Z
authors.lastname: Wen
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Boddicker
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Robin M
authors.initials: RM
authors.lastname: Kaufhold
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Puneet
authors.initials: P
authors.lastname: Khandelwal
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Eberhard
authors.initials: E
authors.lastname: Durr
authors.affiliation: Translational Molecular Biomarkers (Rahway, NJ), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Qiu
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Bob J
authors.initials: BJ
authors.lastname: Lucas
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Debbie D
authors.initials: DD
authors.lastname: Nahas
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: James C
authors.initials: JC
authors.lastname: Cook
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Sinoeun
authors.initials: S
authors.lastname: Touch
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Julie M
authors.initials: JM
authors.lastname: Skinner
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Amy S
authors.initials: AS
authors.lastname: Espeseth
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA.
authors.forename: Craig T
authors.initials: CT
authors.lastname: Przysiecki
authors.affiliation: Infectious Diseases and Vaccines Discovery (West Point, PA), MRL, Merck & Co., Inc, Kenilworth, NJ, USA. lan_zhang2@merck.com.
authors.forename: Lan
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D002478:Cells, Cultured; D002690:Chlamydia Infections / Q000517:prevention & control; D002692:Chlamydia trachomatis / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005260:Female; D000071497:Immunogenicity, Vaccine; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D022223:Vaccines, Subunit / Q000276:immunology; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic
keywords: Chlamydia trachomatis; Immunogenicity; MOMP; Outer membrane expression