Patent application EP19170231A (EP), filed 2019-04-18
Publication number: EP3725371A1
Title: METHOD FOR THE PRODUCTION OF OUTER MEMBRANE VESICLES AND IMMUNOGENIC COMPOSITIONS THEREOF
Abstract: [0001]    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 such bacteria and immunogenic compositions thereof.
IPC codes: A61K  39/   095            A I
A61K  39/   104            A I
A61K  39/   108            A I
A61K  39/   112            A I
A61P  31/    00            A I
A61P  35/    00            A I
C12N  15/    62            A I
Claims: number: 1
text: The use of a gram-negative bacterium for the preparation of isolated outer membrane vesicles (OMVs), wherein said bacterium:
number: 2
text: The use of a bacterium according to claim 1, wherein at least one, preferably at least 5, more preferably at least 10 and even more preferably all of the following genes are inactivated in addition to the ompA gene:
number: 3
text: The use of a bacterium according to claims 1-2, wherein the following genes do not carry gene-inactivating mutations:
number: 4
text: The use of a bacterium according to claims 1-3, which is of the genus Escherichia, Pseudomonas, Neisseria or Shigella.
number: 5
text: The use of a bacterium according to claim 4, which is E. coli.
number: 6
text: The use of a bacterium according to claims 1-5, wherein the genes are inactivated by one of the following methods: point mutations which create a stop codon in the reading frame; deletions of one or multiple nucleotides which impair protein functions; complete deletion of the gene; or inactivation of the transcription and translation signals.
number: 7
text: The use of a bacterium according to claim 1, wherein said heterologous proteins are localized in the lumen of OMVs or associated to the OMV membrane.
number: 8
text: The use of a bacterium according to claim 7, wherein said heterologous proteins are bacterial, viral, parasitic or cancer proteins.
number: 9
text: An outer membrane vesicle (OMV) isolated from a gram-negative bacterium as defined in claim 1.
number: 10
text: An isolated outer membrane vesicle according to claim 9, carrying heterologous proteins which are localized in the lumen of the OMV or associated to the OMV membrane and which are bacterial, viral, parasitic or cancer proteins.
number: 11
text: An isolated outer membrane vesicle according to claim 9, wherein the gram-negative bacterium is Escherichia coli.
number: 12
text: An immunogenic composition comprising an outer membrane vesicle according to claims 9-11, optionally in combination with pharmaceutically acceptable adjuvants and excipients.
number: 13
text: An outer membrane vesicle according to claims 9-11 or an immunogenic composition according to claim 12, for use in the stimulation of an immune response in a subject in need thereof.
number: 14
text: An outer membrane vesicle or an immunogenic composition for use according to claim 11, wherein said subject is affected by an infectious or tumoral disease.
number: 15
text: A method for preparing bacterial outer membrane vesicles (OMVs) which comprises:
number: 16
text: A method according to claim 15, wherein the OMVs are separated from the culture medium by filtration.
number: 17
text: A method according to claim 16, wherein the OMVs are purified by centrifugation or ultrafiltration.
Office action: search_report, dated 2020-10-21
Rejection reasons: novelty_epc_54
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severity: novelty_destroying
ref_id: - ADAM J. KULP ET AL, "Genome-Wide Assessment of Outer Membrane Vesicle Production in Escherichia coli", PLOS ONE, (20150925), vol. 10, no. 9, doi:10.1371/journal.pone.0139200, page e0139200, XP055624792 [A] 1-17 * the whole document *
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ref_id: - KIM S H ET AL, "Structural modifications of outer membrane vesicles to refine them as vaccine delivery vehicles", BIOCHIMICA ET BIOPHYSICA ACTA (BBA) - BIOMEMBRANES, ELSEVIER, AMSTERDAM, NL, vol. 1788, no. 10, doi:10.1016/J.BBAMEM.2009.08.001, ISSN 0005-2736, (20091001), pages 2150 - 2159, (20090818), XP026684863 [A] 1-17 * the whole document *
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ref_id: - GERRITZEN MATTHIAS J H ET AL, "Bioengineering bacterial outer membrane vesicles as vaccine platform", BIOTECHNOLOGY ADVANCES, ELSEVIER PUBLISHING, BARKING, GB, (20170515), vol. 35, no. 5, doi:10.1016/J.BIOTECHADV.2017.05.003, ISSN 0734-9750, pages 565 - 574, XP085118072 [A] 1-17 * the whole document *
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severity: background
ref_id: - BROOKE L. DEATHERAGE ET AL, "Biogenesis of bacterial membrane vesicles", MOLECULAR MICROBIOLOGY, (20090601), vol. 72, no. 6, doi:10.1111/j.1365-2958.2009.06731.x, ISSN 0950-382X, pages 1395 - 1407, XP055015136 [A] 1-17 * the whole document *
ref_type: npl
severity: background
Final disposition: pending (2020-10-21)