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Moraxella (branhamella) catarrhalis antigens

A Moraxella and antibody technology, applied in the direction of bacterial antigen components, bacteria, fungi, etc., can solve the problem of differences in the degree of antibody cross-reactivity

Inactive Publication Date: 2005-02-16
ID BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are differences in the degree of antibody cross-reactivity of proteins between different strains

Method used

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  • Moraxella (branhamella) catarrhalis antigens
  • Moraxella (branhamella) catarrhalis antigens
  • Moraxella (branhamella) catarrhalis antigens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0171] This example illustrates the cloning and molecular characteristics of the BVH-MC6 gene and the corresponding polypeptide.

[0172] M. catarrhalis BVH-MC6 (SEQ ID NO: 1) Gene coding region using the following oligomers containing base overhangs for the addition of restriction sites NdeI (CATATG) and XhoI (CTCGAG): DMAR598 (5'-TAAGGATACATATGACGGCCCATAAAGATCG-3'); DMAR599 (5'-TATGCTCGAGGTATACTTTGACTGGCTTATCATGTG -3′). PCR products were purified from agarose gels using the QIAquick Gel Extraction Kit from QIAgen according to the manufacturer's instructions (Chatsworth, CA) and digested with NdeI and XhoI (Amersham Pharmacia Biotech, Inc, Baie d'Urfé, Canada). The pET21b(+) vector (Novagen, Madison, WI) was digested with NdeI and XhoI and purified from agarose gel using the QIAgen (Chatsworth, CA) QIAquick gel extraction kit. The NdeI-XhoI PCR product was ligated into the NdeI-XhoIpET21b(+) expression vector. According to the Simanis method (Hanahan, D.DNA Cloning, 1985, ...

Embodiment 2

[0178] This example illustrates the cloning and molecular characteristics of the BVH-MC7 gene and the corresponding polypeptide.

[0179] M. catarrhalis BVH-MC7 (SEQ ID NO: 3) For the gene coding region, the following oligomers containing base overhangs for adding restriction sites NdeI (CATATG) and XhoI (CTCGAG) were used: DMAR594 and DMAR691, as shown in Table 1. The method for cloning BVH-MC7 into an expression vector and sequencing is similar to Example 1.

[0180] It has been determined that the open reading frame (ORF) encoding BVH-MC7 contains 1179bp, encoding a polypeptide of 392 amino acid residues, its predicted pI is 8.65, and its predicted molecular weight is 41456.50Da. Using Spscan software (Wisconsin Sequence Analysis Package; Genetics Computer Group) to analyze the predicted amino acid residue sequence (SEQ ID NO: 4), it is suggested that there is a signal peptide (MYQRFINTALVAALAVTMAGC) of 21 amino acid residues, between cysteine ​​and glycine residues ends ...

Embodiment 3

[0183] This example illustrates the cloning of M. catarrhalis genes in the CMV plasmid pCMV-GH.

[0184] The Moraxella catarrhalis polypeptide DNA coding region was inserted into the plasmid vector pCMV-GH, located downstream of the human growth hormone (hGH) gene under the transcriptional control of the cytomegalovirus (CMV) promoter (Tang et al., Nature, 1992, 356 :152). The CMV promoter is a non-functional plasmid in E. coli cells, but is activated after administration of the plasmid to eukaryotic cells. This vector also introduces an ampicillin resistance gene.

[0185] BVH-MC6 (SEQ ID NO: 1) and BVH-MC7 were amplified by PCR (DNA Thermal Cycler GeneAmp PCR System 2400, Perkin Elmer, San Jose, CA) from genomic DNA of Moraxella catarrhalis strain ETSU C-2 (SEQ ID NO: 3) The coding region of the gene without the leader peptide region, using the oligonucleotide primers shown in Table 1, which contains the restriction sites BamHI (GGATCC), BglII (AGATCT), SalI (GTCGAC) or B...

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Abstract

The present invention relates to polypeptides of Moraxella (Branhamella) catarrhalis which may be useful for prophylaxis, diagnostic and / or therapy purposes.

Description

field of invention [0001] The present invention relates to polypeptides that may be used for the prevention, diagnosis and / or treatment of Moraxella (Branhamella) catarrhalis infection, in particular Moraxella catarrhalis (Branhamella) Bacteria) peptides. Background of the invention [0002] Moraxella catarrhalis (Branhamella spp.) is a Gram-negative diplococcus that causes respiratory infections in humans. Moraxella catarrhalis is now considered the third most common cause of otitis media in infants and young children, after Streptococcus pneumoniae and Haemophilus influenzae. M. catarrhalis has also been associated with several other types of infections, including sinusitis, persistent cough, acute laryngitis in adults, suppurative keratitis, neonatal conjunctivitis, and invasive disease in immunocompromised hosts. [0003] Approximately 90% of M. catarrhalis strains are resistant to antibiotics (β-lactamase positive) and the high incidence of recurrent otitis media nece...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53A61K38/00A61K39/00A61K39/02A61P11/02A61P11/04A61P11/06A61P27/02A61P27/16A61P31/04C07H21/04C07K14/195C07K14/21C07K19/00C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12N15/31C12N15/74C12P21/02G01N33/569
CPCC07K2319/00C07K14/212A61K39/00A61P11/02A61P11/04A61P11/06A61P27/02A61P27/16A61P31/04
Inventor D·马丁J·哈枚尔B·R·布罗德尔S·利欧克斯G·莱布兰克J·科特尔
Owner ID BIOMEDICAL
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