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Novel immunogenic compositions for the prevention and treatment of meningococcal disease

An immunogenic, Neisseria meningitidis technology, applied in the field of Neisseria ORF2086 protein, which can solve problems such as the absence of mathematical algorithms

Inactive Publication Date: 2010-01-20
WYETH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even with knowledge of the Neisserial genome, identifying new immunogenic composition candidates remains a challenging process due to insufficient mathematical algorithms currently existing

Method used

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  • Novel immunogenic compositions for the prevention and treatment of meningococcal disease
  • Novel immunogenic compositions for the prevention and treatment of meningococcal disease
  • Novel immunogenic compositions for the prevention and treatment of meningococcal disease

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0185] Identification of Neisseria membrane protein extracts capable of eliciting bactericidal antibodies against heterologous strains:

[0186] Referring to Table II below, LOS (Lipo-Oligosaccharide)-depleted outer membrane protein preparations have been shown to elicit bactericidal antibodies. These antibodies are often directed against the PorA of the respective strain. The LOS-depleted outer membrane preparation from serogroup B meningococcal strain 8529 (B:15:P1.7b,3) is unusual in this manner as it unexpectedly elicits bactericidal antibodies against several heterologous strains .

[0187] Form II

[0188] BC activity against SOMP against different strains of Neisseria meningitidis

[0189]

[0190]

[0191]To facilitate the isolation and characterization of antigens responsible for eliciting heterologous bactericidal antibodies, we sought to identify which detergents would optimally extract antigens.

[0192] Strains and culture conditions

[0193] Neisseria ...

example 2

[0237] Cloning of recombinant lipidated P2086 (RLP2086):

[0238] A.) Native leader sequence:

[0239] Source material:

[0240] The ORF 2086 gene was amplified by PCR from a clinical isolate of a serogroup B Neisseria meningitidis strain designated 8529. The serogroup, serotype and serosubtype of this strain are shown in brackets; 8529 (B: 15, P1: 7b, 3). This meningococcal strain was derived from The RIVM (Bilthoven, The Netherlands).

[0241] PCR amplification and cloning strategies:

[0242] Visual inspection of ORF 2086 indicated that this gene has a potential lipoprotein signal sequence. Additional analysis using a proprietary Hidden Markov Model lipoprotein algorithm confirmed that ORF 2086 contains a lipoprotein signal sequence. For recombinant expression of P2086 in a more native-like conformation, oligonucleotide primers were designed to amplify the full-length gene with the lipoprotein signal sequence intact and based on Sanger sequence analysis of the N. menin...

example 3

[0274] Developmental genetics of the non-lipidated mature 2086 protein:

[0275] To further assess the immunogenicity of the 2086 protein, cloning and expression of the non-lipidated form of P2086 was performed.

[0276] PCR gene amplification of ORF 2086:

[0277] The 2086 gene from various stains is available as in PCT / US02 / 32369 (published as WO 03 / 063766 on Aug. 7, 2003) and PCT / US04 / 11901 (published as WO 04 / 094596 on Nov. 4, 2004) the primers to amplify.

[0278] Features of these primers include a synthetic BglII restriction site in each primer, a synthetic NdeI restriction site in compound nos. 6406 and 6474, and stop codons present in all three reading frames in compound nos. 5135 and 6605. Primer numbers 6406 and 6474 amplify the 2086 gene with ATG (Met) fused to the second amino terminal codon (ACG), which represents a single amino acid substitution of the mature 2086 polypeptide (replacement TGC Cys).

[0279] The PCR cloning vector was TOPO-PCR2.1, Invitrogen, ...

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Abstract

The present invention relates to Neisseria ORF2086 proteins, crossreactive immunogenic proteins which can be isolated from nesserial strains or prepared recombinantly, including immunogenic portions thereof, biological equivalents thereof, antibodies that immunospecifically bind to the foregoing and nucleic acid sequences encoding each of the foregoing, as well as the use of same in immunogenic compositions that are effective against infection by Neisseria meningitidis serogroup B.

Description

technical field [0001] The present invention relates to Neisseria ORF2086 proteins (subgroups A and B) which can be isolated from bacterial strains such as Neisseria species, including Neisseria meningitidis (serogroups A, B, C, D , W-135, X, Y, Z and 29E), strains of Neisseria gonorrhoeae and Neisseria lactamica, and immunogenic portions and / or bioequivalents of said proteins. The present invention also relates to antibodies that immunospecifically bind to said proteins, immunogenic moieties and / or biological equivalents. Furthermore, the present invention relates to isolated polynucleotides comprising nucleic acid sequences encoding any of the aforementioned proteins, immunogenic moieties, biological equivalents and / or antibodies. In addition, the present invention relates to immunogenic compositions and their use in the prevention, treatment and / or diagnosis of meningococcal infections caused by N. meningitidis and in particular by N. meningitidis serogroup B Use in menin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K39/00
CPCA61K39/095A61K2039/541C07K14/22A61P31/04A61P37/04
Inventor 加里·W·兹络特尼克
Owner WYETH LLC
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