title: Quantitative proteomics reveals distinct differences in the protein content of outer membrane vesicle vaccines.
abstract: At present, only vaccines containing outer membrane vesicles (OMV) have successfully stopped Neisseria meningitidis serogroup B epidemics. These vaccines however require detergent-extraction to remove endotoxin, which changes immunogenicity and causes production difficulties. To investigate this in more detail, the protein content of detergent-extracted OMV is compared with two detergent-free alternatives. A novel proteomics strategy has been developed that allows quantitative analysis of many biological replicates despite inherent multiplex restrictions of dimethyl labeling. This enables robust statistical analysis of relative protein abundance. The comparison with detergent-extracted OMV reveales that detergent-free OMV are enriched with membrane (lipo)proteins and contain less cytoplasmic proteins due to a milder purification process. These distinct protein profiles are substantiated with serum blot proteomics, confirming enrichment with immunogenic proteins in both detergent-free alternatives. Therefore, the immunogenic protein content of OMV vaccines depends at least partially on the purification process. This study demonstrates that detergent-free OMV have a preferred composition.
authors: Institute for Translational Vaccinology (Intravacc) , Bilthoven, The Netherlands.
authors: Bas
authors: B
authors: van de Waterbeemd
authors: Geert P M
authors: GP
authors: Mommen
authors: Jeroen L A
authors: JL
authors: Pennings
authors: Michel H
authors: MH
authors: Eppink
authors: René H
authors: RH
authors: Wijffels
authors: Leo A
authors: LA
authors: van der Pol
authors: Ad P J M
authors: AP
authors: de Jong
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D005260:Female; D008074:Lipoproteins / Q000032:analysis / Q000737:chemistry; D022401:Meningococcal Vaccines / Q000032:analysis* / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D038541:Neisseria meningitidis, Serogroup B / Q000472:pathogenicity; D040901:Proteomics / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008074:Lipoproteins; D022401:Meningococcal Vaccines