An improved process for producing outer membrane vesicles

An outer membrane vesicle, physiological technology, applied in the field of medical microbiology and vaccinology, can solve the problems of cell death bacteria, lysis and so on

Pending Publication Date: 2020-07-17
INTRAVACC有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the addition of hydrogen peroxide is not feasible for a scalable OMV production method because the addition of hydrogen peroxide to bacterial cultures results in significant cell death and bacterial lysis

Method used

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  • An improved process for producing outer membrane vesicles
  • An improved process for producing outer membrane vesicles
  • An improved process for producing outer membrane vesicles

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Embodiment Construction

[0016] We have unexpectedly found that the spontaneous release of outer membrane vesicles (OMVs) can be triggered directly and reliably using common control parameters of bioreactor culture.

[0017] In a first aspect, the present invention relates to a method of producing spontaneously released bacterial outer membrane vesicles. Preferably, the method comprises the step of: a) culturing the Gram-negative bacterial population comprising stimulating OMV release by applying a DOT higher than the physiological dissolved oxygen tension (DOT) of 30%. The method also preferably comprises the step of: b) recovering the OMVs released in a), wherein preferably the recovering comprises at least removing bacteria from the OMVs.

[0018] As used herein, the term OMV ("outer membrane vesicle") denotes a released outer membrane sphere with a periplasmic content comprising bioactive molecules (toxins, protein, DNA). OMV is also sometimes referred to as "bleb". These vesicles are often inv...

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Abstract

The present invention relates to the fields of medical microbiology and vaccines. In particular the invention relates to a process wherein the spontaneous release of bacterial outer membrane vesicles(OMV) of Gram-negative bacteria is stimulated by application of a dissolved oxygen tension (DOT) that is higher than a physiological DOT. The thus produced OMVs are for use in vaccines. The inventionfurther relates to OMV obtainable by said process, and to a pharmaceutical composition comprising such OMV. The present invention further relates to the use of OMV of the present invention as a medicament in particular for use in a method for eliciting an immune response.

Description

technical field [0001] The invention relates to the fields of medical microbiology and vaccinology. In particular, the present invention relates to a method for producing Gram-negative bacterial spontaneous outer membrane vesicles (OMVs) for use in vaccines, OMVs obtainable by said method, and pharmaceutical compositions comprising such OMVs. The invention also relates to the use of an OMV according to the invention as a medicament, in particular in a method of eliciting an immune response. Background technique [0002] Outer membrane vesicles (OMVs) are naturally produced by Gram-negative bacteria and are involved in pathogenesis, intercellular communication, and stress response (Kulp and Kuehn 2010). Recent studies have shown membrane vesicle formation in Gram-positive bacteria and archaea (Ellen et al. 2009; Rivera et al. 2010). OMVs are spherical nanoparticles, vesicles composed of a phospholipid bilayer containing proteins and lipopolysaccharide (LPS), and the vesicle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04C12P21/02A61K39/095A61K39/02A61K39/108
CPCA61K39/0208A61K39/0258A61K39/095A61K39/099C12P21/02C12N1/20C12P1/04Y02A50/30C12M41/34A61P31/00C12N2509/10A61K2039/55555
Inventor M·J·H·格里岑L·A·范德波尔M·施托克
Owner INTRAVACC有限责任公司
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