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

Wei He, Martina Felderman, Angela C. Evans, Jia Geng, David Homan, Feliza Bourguet, Nicholas O Fischer, Yuanpei Li, Kit Lam, Aleksandr Noy, Li Xing, R. Holland Cheng, Amy Rasley, Craig D. Blanchette, Kurt Kamrud, Nathaniel Wang, Heather Gouvis, Todd C. Peterson, Bolyn Hubby, Matthew A Coleman

Research output: Contribution to journalArticle

2 Citations (Scopus)

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. Longterm 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 membranebound proteins for future vaccine development. This method can be applied to other previously difficult-to-obtain antigens while maintaining full functionality and immunogenicity.

Original languageEnglish (US)
Pages (from-to)15121-15132
Number of pages12
JournalJournal of Biological Chemistry
Volume292
Issue number36
DOIs
StatePublished - 2017

Fingerprint

Chlamydia
Membrane Proteins
Vaccines
Proteins
Chlamydia muridarum
Antigens
Pelvic Inflammatory Disease
Chlamydia Infections
Cell-Free System
Oligodeoxyribonucleotides
Single-Stranded DNA
Chlamydia trachomatis
Public health
Blindness
Humoral Immunity
Sexually Transmitted Diseases
Codon
Solubility
Escherichia coli
Infertility

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development. / He, Wei; Felderman, Martina; Evans, Angela C.; Geng, Jia; Homan, David; Bourguet, Feliza; Fischer, Nicholas O; Li, Yuanpei; Lam, Kit; Noy, Aleksandr; Xing, Li; Cheng, R. Holland; Rasley, Amy; Blanchette, Craig D.; Kamrud, Kurt; Wang, Nathaniel; Gouvis, Heather; Peterson, Todd C.; Hubby, Bolyn; Coleman, Matthew A.

In: Journal of Biological Chemistry, Vol. 292, No. 36, 2017, p. 15121-15132.

Research output: Contribution to journalArticle

He, W, Felderman, M, Evans, AC, Geng, J, Homan, D, Bourguet, F, Fischer, NO, Li, Y, Lam, K, Noy, A, Xing, L, Cheng, RH, Rasley, A, Blanchette, CD, Kamrud, K, Wang, N, Gouvis, H, Peterson, TC, Hubby, B & Coleman, MA 2017, 'Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development', Journal of Biological Chemistry, vol. 292, no. 36, pp. 15121-15132. https://doi.org/10.1074/jbc.M117.784561
He, Wei ; Felderman, Martina ; Evans, Angela C. ; Geng, Jia ; Homan, David ; Bourguet, Feliza ; Fischer, Nicholas O ; Li, Yuanpei ; Lam, Kit ; Noy, Aleksandr ; Xing, Li ; Cheng, R. Holland ; Rasley, Amy ; Blanchette, Craig D. ; Kamrud, Kurt ; Wang, Nathaniel ; Gouvis, Heather ; Peterson, Todd C. ; Hubby, Bolyn ; Coleman, Matthew A. / Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 36. pp. 15121-15132.
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AU - Homan, David

AU - Bourguet, Feliza

AU - Fischer, Nicholas O

AU - Li, Yuanpei

AU - Lam, Kit

AU - Noy, Aleksandr

AU - Xing, Li

AU - Cheng, R. Holland

AU - Rasley, Amy

AU - Blanchette, Craig D.

AU - Kamrud, Kurt

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