Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA
- Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Merced, CA (United States). School of Natural Sciences
- Univ. of California, Davis, CA (United States). Dept. of Biochemistry and Molecular Medicine
- Univ. of California, Davis, CA (United States). Dept. of Molecular and Cellular Biology
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Davis, CA (United States). School of Medicine, Radiation Oncology
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 the 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.
- Research Organization:
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
- Sponsoring Organization:
- USDOE; National Institutes of Health (NIH); Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
- Grant/Contract Number:
- AC52-07NA27344; R01CA115483; R01CA140449; R21 RAI120925A
- OSTI ID:
- 1438621
- Report Number(s):
- LLNL-JRNL-747580
- Journal Information:
- Journal of Biological Chemistry, Vol. 292, Issue 36; ISSN 0021-9258
- Publisher:
- American Society for Biochemistry and Molecular BiologyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
High-yield production of “difficult-to-express” proteins in a continuous exchange cell-free system based on CHO cell lysates
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journal | September 2017 |
Cell-Free Co-Translational Approaches for Producing Mammalian Receptors: Expanding the Cell-Free Expression Toolbox Using Nanolipoproteins
|
journal | July 2019 |
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