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1. WO2004019977 - IMPROVED BACTERIAL OUTER MEMBRANE VESICLES

Publication Number WO/2004/019977
Publication Date 11.03.2004
International Application No. PCT/IB2003/004293
International Filing Date 01.09.2003
Chapter 2 Demand Filed 30.03.2004
IPC
A61K 9/00 2006.01
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
9Medicinal preparations characterised by special physical form
A61K 39/095 2006.01
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39Medicinal preparations containing antigens or antibodies
02Bacterial antigens
095Neisseria
C12N 1/06 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
06Lysis of microorganisms
CPC
A61K 2039/55555
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39Medicinal preparations containing antigens or antibodies
555characterised by a specific combination antigen/adjuvant
55511Organic adjuvants
55555Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
A61K 39/095
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39Medicinal preparations containing antigens or antibodies
02Bacterial antigens
095Neisseria
A61P 31/04
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
31Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
04Antibacterial agents
A61P 37/04
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
37Drugs for immunological or allergic disorders
02Immunomodulators
04Immunostimulants
C07K 14/22
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195from bacteria
22from Neisseriaceae (F)
C12N 1/06
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof
06Lysis of microorganisms
Applicants
  • CHIRON SRL [IT]/[IT] (AllExceptUS)
  • PIZZA, Mariagrazia [IT]/[IT] (UsOnly)
  • SERRUTO, Davide [IT]/[IT] (UsOnly)
  • RAPPUOLI, Rini [IT]/[IT] (UsOnly)
Inventors
  • PIZZA, Mariagrazia
  • SERRUTO, Davide
  • RAPPUOLI, Rini
Agents
  • MARSHALL, Cameron, John
Priority Data
0220194.530.08.2002GB
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) IMPROVED BACTERIAL OUTER MEMBRANE VESICLES
(FR) AMELIORATIONS APPORTEES A DES VESICULES DE MEMBRANE EXTERNE BACTERIENNES
Abstract
(EN)
Existing methods of meningococcal OMV preparation involve the use of detergent during disruption of the bacterial membrane. According to the invention, membrane disruption is performed substantially in the absence of detergent. The resulting OMVs which retain important bacterial immunogenic components, particularly (i) the protective NspA surface protein, (ü) protein NMB2132 and (iii) protein NMB 1870.A Typical process involves the following steps: (a) treating bacterial cells in the substantial absence of detergent; (b) centrifuging the composition from step (a) to separate the outer membrane vesicles from treated cells and cell debris, and collecting the supernatant; (c) performing a high speed centrifugation of the supernatant from step (b) and collecting the outer membrane vesicles in a pellet; (d) re-dispersing the pellet from step (c) in a buffer; (e) performing a second high speed centrifugation in accordance with step (c), collecting the outer membrane vesicles in a pellet; (f) re-dispersing the pellet from step (e) in an aqueous medium.
(FR)
Les procédés existants pour la préparation de vésicules de membrane externe méningococciques impliquent l'utilisation de détergent lors de la rupture de la membrane bactérienne. Selon l'invention, cette rupture est réalisée pratiquement sans détergent. L'invention concerne également les vésicules de membrane externe obtenues, qui renferment des composants immunogènes bactériens importants, en particulier (i) la protéine de surface NspA protectrice, (ii) la protéine NMB2132 et (iii) la protéine NMB 1870. Un procédé typique selon l'invention comprend les étapes suivantes : (a) traitement des cellules bactériennes pratiquement sans détergent; (b) centrifugation de la composition obtenue à l'étape (a) pour séparer les vésicules de membrane externe des cellules traitées et des débris de cellules, et collecte du surnageant; (c) centrifugation à vitesse élevée du surnageant obtenu à l'étape (b) et collecte des vésicules de membrane extérieure dans une pastille; (d) redispersion de la pastille obtenue à l'étape (c) dans un tampon; (e) deuxième centrifugation à vitesse élevée selon l'étape (c), collecte des vésicules de membrane externe dans une pastille; (f) redispersion de la pastille obtenue à l'étape (e) dans un milieu aqueux.
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