Streptococcus antigens

a technology of streptococcus pneumoniae and antigens, which is applied in the field of antibodies, epitopes and antibodies, can solve the problems of many deaths, failure of existing vaccines and capsular conjugate vaccines, and the presence of resistant pneumococcal organisms,

Inactive Publication Date: 2006-08-10
ID BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even with appropriate antibiotic therapy, pneumococcal infections still result in many deaths.
Although the advent of antimicrobial drugs has reduced the overall mortality from pneumococcal disease, the presence of resistant pneumococcal organisms has become a major problem in the world today.
In particular, the failure of existing vaccines and capsular conjugate vaccines currently in development to protect young children against all serotypes spurres evaluation of other S. pneumoniae components.
Although immunogenicity of capsular polysaccharides can be improved, serotype specificity will still represent a major limitation of polysaccharide-based vaccines.
However, no biological activity of these polypeptides is reported.

Method used

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Examples

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example 1

[0166] This example describes the bacterial strains, plasmids, PCR primers, recombinant proteins and hybridoma antibodies used herein.

[0167]S. pneumoniae SP64 (serogroup 6) and SP63 (serogroup 9) clinical isolates were provided by the Laboratoire de la Santé Publique du Québec, Sainte-Anne-de-Bellevue; Rx1 strain, a nonencapsulated derivative of the type 2 strain D39 and the type 3 strain WU2 were provided by David E. Briles from University of Alabama, Birmingham and the type 3 clinical isolate P4241 was provided by the Centre de Recherche en Infectiologie du Centre Hospitalier de l'Universite Laval, Sainte-Foy. E. coli strains DH5α (Gibco BRL, Gaithesburg, Md.); AD494 (λDE3) (Novagen, Madison, Wis.) and BL21 (λDE3) (Novagen) as well as plasmid superlinker pSL301 vector (Invitrogen, San Diego, Calif.); PCMV-GH vector (gift from Dr. Stephen A. Johnston, Department for Biochemistry, University of Texas, Dallas, Tex.); pET32 and pET21 (Novagen) and pURV22.HIS expression vectors (FIG. ...

example 2

[0173] This example describes the identification of peptide domains carrying target epitopes using Mabs and recombinant truncated proteins described in example 1.

[0174] Hybridomas were tested by ELISA against truncated BVH-3, BVH-11 or BVH-11-2 gene products in order to characterize the epitopes recognized by the Mabs. The truncated gene products were generated from S. pneumoniae SP64 strain except for BVH-3-Sp63 which was generated from S. pneumoniae SP63 strain. As a positive control, the reactivity of each antibody was examined with full-length BVH-3, BVH-11 or BVH-11-2 recombinant proteins. In some cases, the Mab reactivity was evaluated by Western immunoblotting after separation of the gene product by SDS-PAGE and transfer on nitrocellulose paper. The reactivities observed is set forth in the following Table 3 and Table 4.

TABLE 3ELISA reactivity of BVH-3-reactive Mabs with a panel of eleven BVH-3 geneproducts and the BVH-11M moleculeMabsGene products tested(IgGBVH-BVH-BVH-BV...

example 3

[0179] This example describes the construction of BVH-3 and BVH-11-2 gene libraries for the mapping of epitopes.

[0180] BVH-3 and BVH-11-2 gene libraries were constructed using recombinant PCMV-GH and PSL301 plasmid DNA containing respectively, BVH-3 gene sequence spanning nucleotides 1837 to 4909 (SEQ ID NO: 2) or BVH-11-2 gene sequence spanning nucleotides 172 to 2630 (SEQ ID NO: 5) and the Novatope® library construction and screening system (Novagen). The recombinant plasmids containing BVH-3 or BVH-11-2 gene fragment were purified using QIAgen kit (Chatsworth, Calif.) and digested with the restriction enzymes BglII and XbaI respectively. The resulting BglII-XbaI DNA fragments were purified using the QIAquick gel extraction kit from QIAgen and digested with Dnase I for the generation of randomly cleaved DNA. DNA fragments of 50 to 200 bp were purified, treated with T4 DNA polymerase to blunt the target DNA ends and add a single 3′dA residue, and ligated into pSCREEN-T-Vector (Nov...

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Abstract

Streptococcus polypeptides and polynucleotides encoding them are disclosed. Said polypeptides may be useful vaccine components for the prophylaxis or therapy of streptococcus infection in animals. Also disclosed are recombinant methods of producing the protein antigens as well as diagnostic assays for detecting streptococcus bacterial infection.

Description

[0001] This application claims the benefit of U.S. provisional application 60 / 212,683 filed Jun. 20, 2000 which is herein incorporated by reference.FIELD OF THE INVENTION [0002] The present invention is related to antigens, epitopes and antibodies directed to these epitopes, more particularly polypeptide antigens of streptococcus pneumoniae pathogen which may be useful for prophylaxis, diagnostic or treatment of streptococcal infection. BACKGROUND OF THE INVENTION [0003]S. pneumoniae is an important agent of disease in man especially among infants, the elderly and immunocompromised persons. It is a bacterium frequently isolated from patients with invasive diseases such as bacteraemia / septicaemia, pneumonia, meningitis with high morbidity and mortality throughout the world. Even with appropriate antibiotic therapy, pneumococcal infections still result in many deaths. Although the advent of antimicrobial drugs has reduced the overall mortality from pneumococcal disease, the presence o...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/02C07H21/04C12N1/20C07K14/315C12N15/74A61K39/00C12N1/21
CPCA61K39/00C07K14/315
Inventor HAMEL, JOSEEOUELLET, CATHERINECHARLAND, NATHALIEMARTIN, DENISBRODEUR, BERNARD
Owner ID BIOMEDICAL
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