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Title: Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development

Chlamydia is a prevalent sexually transmitted disease that infects more than 100 million people worldwide. Although most individuals infected with Chlamydia trachomatis are initially asymptomatic, symptoms can arise if left undiagnosed. Long-term infection can result in debilitating conditions such as pelvic inflammatory disease, infertility, and blindness. Chlamydia infection, therefore, constitutes a significant public health threat, underscoring the need for a Chlamydia-specific vaccine. Chlamydia strains express a major outer-membrane protein (MOMP) that has been shown to be an effective vaccine antigen. However, approaches to produce a functional recombinant MOMP protein for vaccine development are limited by poor solubility, low yield, and protein misfolding. For this study, we used an Escherichia coli-based cell-free system to express a MOMP protein from the mouse-specific species Chlamydia muridarum (MoPn-MOMP or mMOMP). The codon-optimized mMOMP gene was co-translated with Δ49apolipoprotein A1 (Δ49ApoA1), a truncated version of mouse ApoA1 in which the N-terminal 49 amino acids were removed. This co-translation process produced mMOMP supported within a telodendrimer nanolipoprotein particle (mMOMP–tNLP). The cell-free expressed mMOMP–tNLPs contain mMOMP multimers similar to the native MOMP protein. This cell-free process produced on average 1.5 mg of purified, water-soluble mMOMP–tNLP complex in a 1-ml cell-free reaction. The mMOMP–tNLP particle also accommodated themore » co-localization of CpG oligodeoxynucleotide 1826, a single-stranded synthetic DNA adjuvant, eliciting an enhanced humoral immune response in vaccinated mice. Using our mMOMP–tNLP formulation, we demonstrate a unique approach to solubilizing and administering membrane-bound proteins for future vaccine development. This method can be applied to other previously difficult-to-obtain antigens while maintaining full functionality and immunogenicity.« less
 [1] ;  [2] ;  [1] ;  [3] ;  [1] ;  [1] ;  [1] ;  [4] ;  [4] ;  [3] ;  [5] ;  [5] ;  [1] ;  [1] ;  [2] ;  [2] ;  [2] ;  [2] ;  [2] ;  [6]
  1. Lawrence Livermore National Laboratory (LLNL), Livermore, CA
  2. Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
  3. Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Merced, CA (United States). School of Natural Sciences
  4. Univ. of California, Davis, CA (United States). Dept. of Biochemistry and Molecular Medicine
  5. Univ. of California, Davis, CA (United States). Dept. of Molecular and Cellular Biology
  6. Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Davis, CA (United States). School of Medicine, Radiation Oncology
Publication Date:
Report Number(s):
Journal ID: ISSN 0021-9258
Grant/Contract Number:
AC52-07NA27344; R01CA115483; R01CA140449; R21 RAI120925A
Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 292; Journal Issue: 36; Journal ID: ISSN 0021-9258
American Society for Biochemistry and Molecular Biology
Research Org:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org:
USDOE; National Institutes of Health (NIH); Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
Country of Publication:
United States
60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; apolipoprotein; Chlamydia; membrane protein; nanotechnology; oligomer; cell-free expression; major outer membrane protein; nanolipoproteins; telodendrimer
OSTI Identifier: