Meningococcal Outer Membrane Vesicle Production

Outer membrane vesicles were provided by Janssen Vaccines and Prevention B.V. Briefly, OMVs were produced from N. meningitidis H44/76 (B:P1.7,16;F3-3) WT or derivatives. nOMVs were produced from a H44/76 ΔRΔL (15 (link), 16 (link), 27 (link)), resulting in a pentaacylated meningococcal LPS from the lpxL1 mutant N. meningitidis strain, lacking the secondary C12:0 acyl chain at the 2′-position of the lipid A. Detergent-extracted OMVs were produced from the WT strain or the ΔRΔL mutant (28 ). Adsorption to aluminum hydroxide (Sigma-Aldrich) was performed, as described elsewhere (29 (link)). Before use, OMVs were diluted in 10 mM Tris pH 7.4/3% (w/v) sucrose to 50 μg total protein/ml. OMV size was determined by dynamic light scattering (DLS) using the Zetasizer nanoseries (Malvern Nano-ZS, l 1/4 532 nm, Westborough, MA, USA).

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Dowling D.J., Sanders H., Cheng W.K., Joshi S., Brightman S., Bergelson I., Pietrasanta C., van Haren S.D., van Amsterdam S., Fernandez J., van den Dobbelsteen G.P, & Levy O. (2016). A Meningococcal Outer Membrane Vesicle Vaccine Incorporating Genetically Attenuated Endotoxin Dissociates Inflammation from Immunogenicity. Frontiers in Immunology, 7, 562.

Publication 2016

aluminum hydroxide

Adsorption Aluminum hydroxide Derivatives Detergent Lipid a Membrane Meningitidis n Meningococcal Protein Strain Sucrose Tris Vaccines

Corresponding Organization : Boston Children's Hospital

Other organizations : Janssen (Netherlands), Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Janssen (United States)

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Variable analysis

independent variables

Strain of N. meningitidis (wild-type vs. ΔRΔL mutant)
Presence or absence of detergent extraction of OMVs

dependent variables

OMV size determined by dynamic light scattering

control variables

Tris buffer pH 7.4 with 3% (w/v) sucrose used to dilute OMVs before analysis
Adsorption of OMVs to aluminum hydroxide

controls

Positive control: Wild-type N. meningitidis H44/76 strain
Negative control: ΔRΔL mutant N. meningitidis H44/76 strain

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