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Meningococcus adhesins nada, app and orf 40

a technology of meningococcus and adhesins, applied in the field of biochemistry, can solve the problems that do not reveal everything about this pathogen, and achieve the effect of preventing the attachment of a neisserial cell

Inactive Publication Date: 2010-09-02
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent text describes a method for purifying a protein called App, which is found in the meningitis-causing bacteria N. meningitidis. The method involves expressing the App protein in a non-Neisserial host cell, such as E. coli, and then purifying the processed version of the protein from the culture medium. The purified protein can be used for various applications, such as research and development of new treatments for meningitis. The invention also provides a method for expressing and purifying another protein called NadA, which is also found in N. meningitidis. The method involves similar steps and techniques. Overall, the invention provides a way to produce and utilize these important proteins for research and development purposes.

Problems solved by technology

Sequence data alone, however, does not reveal everything about this pathogen.

Method used

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  • Meningococcus adhesins nada, app and orf 40
  • Meningococcus adhesins nada, app and orf 40
  • Meningococcus adhesins nada, app and orf 40

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Experimental program
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Embodiment Construction

NadA Homology

[0195]NadA shows homology to (a) YadA of enteropathogenic Yersinia, a non-pilus associated adhesin implicated in virulence [Cornelis (1998) Microbiol. Mol. Biol. Rev. 62:1315-1352.] and (b) UspA2 of Moraxella catarrhalis, a protein involved in serum resistance and a protective antigen [Chen et al. (1999) Infect. Immun. 67:1310-1316.]. Sequence similarity is mainly clustered in the carboxyl terminal region (56-63% identity in the last 70 amino acids). Outside this region the level of identity drops to 23-25%.

[0196]YadA and UspA2 have been identified as adhesins [Hoiczyk et al. (2000) EMBO J 19:5989-5999]. Both proteins form very stable and difficult-to-dissociate high molecular weight oligomers (150-200 kDa) anchored to the outer membrane. NadA has also been found to form very stable high molecular weight aggregates on the outer membrane of meningococcus.

[0197]The amino acid sequence of NadA was analysed [Nielsen et al. (1997) Protein Engineering 10:1-6; Levin & Garner (...

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Abstract

NadA, App and ORF40 function as adhesins in N. meningitidis. Adhesion can be modulated by targeting these three proteins. NadA allelic variants are disclosed. Autoproteolytic cleavage of App is disclosed, as is removal of the activity by mutagenesis. App is processed and secreted into culture medium when expressed in E. coli. Mature App proteins are disclosed. Knockout mutants are disclosed. Vesicles from non-Neisserial hosts with heterologous adhesin expression are disclosed.

Description

[0001]All documents cited herein are incorporated by reference in their entirety.TECHNICAL FIELD[0002]This invention is in the field of biochemistry and, in particular, the biochemistry of the pathogenic bacteria in the genus Neisseria (e.g. N. meningitidis and N. gonorrhoeae).BACKGROUND ART[0003]International patent applications WO99 / 24578, WO99 / 36544, WO99 / 57280 and WO00 / 22430 disclose proteins from Neisseria meningitidis and Neisseria gonorrhoeae. The complete genome sequence of serogroup B N. meningitidis has been published [Tettelin et al. (2000) Science 287:1809-1815] and has been subjected to analysis in order to identify vaccine antigens [Pizza et al. (2000) Science 287:1816-1820]. Approaches to expression of the proteins are disclosed in WO01 / 64922. The complete genome sequence of serogroup A N. meningitidis is also known [Parkhill et al. (2000) Nature 404:502-506].[0004]Sequence data alone, however, does not reveal everything about this pathogen. Objects of the present inv...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395C07K16/00C12N1/20A61P31/04A61K39/00C12N15/09A61K39/095C07K14/22C12N1/21C12P21/00C12Q1/02C12R1/36
CPCA61K39/00C07K14/22A61K2039/53A61P31/04
Inventor ARICO, MARIACOMANDUCCI, MAURIZIO
Owner NOVARTIS AG
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