Cell-free Scaled Production and Adjuvant Addition to a Recombinant Major Outer Membrane Protein from Chlamydia muridarum for Vaccine Development
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
- University of California, Irvine, CA (United States)
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States); University of California, Davis, CA (United States)
Subunit vaccines offer advantages over more traditional inactivated or attenuated whole-cell-derived vaccines in safety, stability, and standard manufacturing. To achieve an effective protein-based subunit vaccine, the protein antigen often needs to adopt a native-like conformation. This is particularly important for pathogensurface antigens that are membrane-bound proteins. Cell-free methods have been successfully used to produce correctly folded functional membrane protein through the co-translation of nanolipoprotein particles (NLPs), commonly known as nanodiscs. This strategy can be used to produce subunit vaccines consisting of membrane proteins in a lipid-bound environment. However, cell-free protein production is often limited to small scale (<1 mL). The amount of protein produced in small-scale production runs is usually sufficient for biochemical and biophysical studies. However, the cell-free process needs to be scaled up, optimized, and carefully tested to obtain enough protein for vaccine studies in animal models. Other processes involved in vaccine production, such as purification, adjuvant addition, and lyophilization, need to be optimized in parallel. This paper reports the development of a scaled-up protocol to express, purify, and formulate a membrane-bound protein subunit vaccine
- Research Organization:
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
- Sponsoring Organization:
- USDOE National Nuclear Security Administration (NNSA); National Institute of Allergy and Infectious Diseases (NIAID)
- Grant/Contract Number:
- AC52-07NA27344; R21 AI20925; U19 AI144184
- OSTI ID:
- 1959448
- Report Number(s):
- LLNL-JRNL-822525; 1035047
- Journal Information:
- Journal of Visualized Experiments, Vol. vol. 181, Issue 181; ISSN 1940-087X
- Publisher:
- MyJoVE Corp.Copyright Statement
- Country of Publication:
- United States
- Language:
- English
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