1. Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development
- Author
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He, Wei, Felderman, Martina, Evans, Angela C, Geng, Jia, Homan, David, Bourguet, Feliza, Fischer, Nicholas O, Li, Yuanpei, Lam, Kit S, Noy, Aleksandr, Xing, Li, Cheng, R Holland, Rasley, Amy, Blanchette, Craig D, Kamrud, Kurt, Wang, Nathaniel, Gouvis, Heather, Peterson, Todd C, Hubby, Bolyn, and Coleman, Matthew A
- Subjects
Medical Microbiology ,Biomedical and Clinical Sciences ,Clinical Sciences ,Sexually Transmitted Infections ,Nanotechnology ,Prevention ,Immunization ,Vaccine Related ,Biotechnology ,Infectious Diseases ,Bioengineering ,Contraception/Reproduction ,Emerging Infectious Diseases ,Prevention of disease and conditions ,and promotion of well-being ,3.4 Vaccines ,Infection ,Good Health and Well Being ,Animals ,Bacterial Outer Membrane Proteins ,Bacterial Vaccines ,Base Sequence ,Cell-Free System ,Chlamydia Infections ,Chlamydia muridarum ,Female ,Mice ,Mice ,Inbred BALB C ,Chlamydia ,apolipoprotein ,cell-free expression ,major outer membrane protein ,membrane protein ,nanolipoproteins ,nanotechnology ,oligomer ,telodendrimer ,Chemical Sciences ,Biological Sciences ,Medical and Health Sciences ,Biochemistry & Molecular Biology ,Biological sciences ,Biomedical and clinical sciences ,Chemical sciences - Abstract
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. Here, 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.
- Published
- 2017