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US 12,551,544 B2Granted
Method for the production of outer membrane vesicles and immunogenic compositions thereof
AssigneeBiomvis Srl
InventorsGuido Grandi, Alberto Grandi, Laura Fantappie', Ilaria Zanella, Enrico Koenig, Assunta Gagliardi
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Abstract
The invention relates to gram-negative bacteria carrying gene-inactivating mutations that cause deletion of proteins belonging to the OMV proteome, to Outer Membrane Vesicles (OMVs) produced by these bacteria and immunogenic compositions thereof.
Claims12
A method for preparing isolated bacterial outer membrane vesicles (OMVs) expressing heterologous proteins, which comprises:
- (i) culturing a gram-negative bacterium in conditions suitable for vesiculation, wherein said bacterium is E. coli and wherein said bacterium:
- (a) carries gene-inactivating mutations at the ompA gene and at least 5 of the following genes which encode proteins naturally present in the OVMs of E. coli:
- ybis, ais, eco, glpQ, mltA, proX, ydcL, glnH, efeO, bglX, agp, ygdI, yncD, slp, artI, yiaD, ompX, borD, yhiJ, emtA, fecA, nmpC, fhuA, hisJ, lamB, malE, malM, ygiW, cirA, fepA, loiP, yjeI, ecnB, rcsF, phoE, oppA, fkpA, ybaY, tsx, yggE, osmE, ygdR, yceI, bhsA, nlpE, pldA, yghJ, ydeN, ushA, mdoD, treA, bcsC, ftsP, ptrA, fadL, artJ, mlaA;
- (b) expresses heterologous proteins in the OMVs
- (ii) separating the OMVs from the bacterial culture,
- and optionally
- (iii) purifying the OMVs.
4 more elements
Dependencies are inferred from the claim text.