Abstract
Outer membrane vesicles (OMV) represent a promising platform for the development of vaccines against bacterial pathogens. More recently, bacteria have been genetically modified to increase OMV yield and modulate the design of resulting particles, also named generalized modules for membrane antigens (GMMA). OMV/GMMA resemble the bacterial surface of the pathogen, where key antigens to elicit a protective immune response are and contain pathogen-associated molecular patterns (e.g., lipopolysaccharides, lipoproteins) conferring self-adjuvanticity. On the other hand, OMV/GMMA are quite complex molecules and a comprehensive panel of analytical methods is needed to ensure quality, consistency of manufacture and to follow their stability over time. Here, we describe several procedures that can be used for OMV/GMMA characterization as particles and for analysis of key antigens displayed on their surface.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Micoli F, MacLennan CA (2020) Outer membrane vesicle vaccines. Semin Immunol 50:101433. https://doi.org/10.1016/j.smim.2020.101433
Ellis TN, Kuehn MJ (2010) Virulence and immunomodulatory roles of bacterial outer membrane vesicles. Microbiol Mol Biol Rev 74(1):81–94. https://doi.org/10.1128/mmbr.00031-09
Gerke C, Colucci AM, Giannelli C et al (2015) Production of a Shigella sonnei vaccine based on generalized modules for membrane antigens (GMMA), 1790GAHB. PLoS One 10(8):e0134478. https://doi.org/10.1371/journal.pone.0134478
van de Waterbeemd B, Streefland M, van der Ley P et al (2010) Improved OMV vaccine against Neisseria meningitidis using genetically engineered strains and a detergent-free purification process. Vaccine 28(30):4810–4816. https://doi.org/10.1016/j.vaccine.2010.04.082
Mancini F, Rossi O, Necchi F, Micoli F (2020) OMV vaccines and the role of TLR agonists in immune response. Int J Mol Sci 21(12):4416. https://doi.org/10.3390/ijms21124416
van der Pol L, Stork M, van der Ley P (2015) Outer membrane vesicles as platform vaccine technology. Biotechnol J 10(11):1689–1706. https://doi.org/10.1002/biot.201400395
Kis Z, Shattock R, Shah N, Kontoravdi C (2019) Emerging technologies for low-cost, rapid vaccine manufacture. Biotechnol J 14(1):e1800376. https://doi.org/10.1002/biot.201800376
Li M, Zhou H, Yang C et al (2020) Bacterial outer membrane vesicles as a platform for biomedical applications: an update. J Control Release 323:253–268. https://doi.org/10.1016/j.jconrel.2020.04.031
De Benedetto G, Cescutti P, Giannelli C, Rizzo R, Micoli F (2017) Multiple techniques for size determination of generalized modules for membrane antigens from Salmonella typhimurium and Salmonella enteritidis. ACS Omega 2(11):8282–8289. https://doi.org/10.1021/acsomega.7b01173
Tani C, Stella M, Donnarumma D et al (2014) Quantification by LC-MS(E) of outer membrane vesicle proteins of the Bexsero® vaccine. Vaccine 32(11):1273–1279. https://doi.org/10.1016/j.vaccine.2014.01.011
Maggiore L, Yu L, Omasits U et al (2016) Quantitative proteomic analysis of Shigella flexneri and Shigella sonnei generalized modules for membrane antigens (GMMA) reveals highly pure preparations. Int J Med Microbiol 306(2):99–108. https://doi.org/10.1016/j.ijmm.2015.12.003
Micoli F, Alfini R, Di Benedetto R et al (2020) GMMA is a versatile platform to design effective multivalent combination vaccines. Vaccine 8(3):540. https://doi.org/10.3390/vaccines8030540
Giannelli C, Raso MM, Palmieri E et al (2020) Development of a specific and sensitive HPAEC-PAD method for quantification of vi polysaccharide applicable to other polysaccharides containing amino uronic acids. Anal Chem 92(9):6304–6311. https://doi.org/10.1021/acs.analchem.9b05107
De Benedetto G, Alfini R, Cescutti P et al (2017) Characterization of O-antigen delivered by generalized modules for membrane antigens (GMMA) vaccine candidates against nontyphoidal Salmonella. Vaccine 35(3):419–426. https://doi.org/10.1016/j.vaccine.2016.11.089
Rossi O, Aruta MG, Acquaviva A et al (2020) Characterization of competitive ELISA and formulated alhydrogel competitive ELISA (FAcE) for direct quantification of active ingredients in GMMA-based vaccines. Methods Protoc 3(3):62. https://doi.org/10.3390/mps3030062
Rossi O, Caboni M, Negrea A et al (2016) Toll-like receptor activation by generalized modules for membrane antigens from lipid A mutants of Salmonella enterica serovars typhimurium and enteritidis. Clin Vaccine Immunol 23(4):304–314. https://doi.org/10.1128/CVI.00023-16
Micoli F, Ravenscroft N, Cescutti P et al (2014) Structural analysis of O-polysaccharide chains extracted from different Salmonella typhimurium strains. Carbohydr Res 385:1–8. https://doi.org/10.1016/j.carres.2013.12.003
Biagini M, Spinsanti M, De Angelis G et al (2016) Expression of factor H binding protein in meningococcal strains can vary at least 15-fold and is genetically determined. Proc Natl Acad Sci U S A 113(10):2714–2719. https://doi.org/10.1073/pnas.1521142113
Raso MM, Gasperini G, Alfini R et al (2020) GMMA and glycoconjugate approaches compared in mice for the development of a vaccine against Shigella flexneri serotype 6. Vaccine 8(2):160. https://doi.org/10.3390/vaccines8020160
MacLennan CA, Martin LB, Micoli F (2014) Vaccines against invasive Salmonella disease: current status and future directions. Hum Vaccin Immunother 10(6):1478–1493. https://doi.org/10.4161/hv.29054
Mani S, Wierzba T, Walker RI (2016) Status of vaccine research and development for Shigella. Vaccine 34(26):2887–2894. https://doi.org/10.1016/j.vaccine.2016.02.075
Lyngby J, Olsen LH, Eidem T, Lundanes E, Jantzen E (2002) Quantification of lipopolysaccharides in outer membrane vesicle vaccines against meningococcal disease. High-performance liquid chromatographic determination of the constituent 3-hydroxy-lauric acid. Biologicals 30(1):7–13. https://doi.org/10.1006/biol.2001.0285
Macgee J, Doudoroff M (1954) A new phosphorylated intermediate in glucose oxidation. J Biol Chem 210(2):617–626
Acknowledgments
All authors were involved in drafting the book chapter and approved the final version. The development of this article was sponsored by GlaxoSmithKline Biologicals SA. The authors declare the following interests: FM, CG, and RAL are employees of the GSK group of companies.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Micoli, F., Alfini, R., Giannelli, C. (2022). Methods for Assessment of OMV/GMMA Quality and Stability. In: Bidmos, F., Bossé, J., Langford, P. (eds) Bacterial Vaccines. Methods in Molecular Biology, vol 2414. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1900-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1900-1_14
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1899-8
Online ISBN: 978-1-0716-1900-1
eBook Packages: Springer Protocols