Pneumococcus polysaccharide protein coupling vaccine and its preparing method

A pneumococcal polysaccharide and protein coupling technology, applied in the field of biomedicine, can solve the problems of T cell memory, low titer, toxicity of pneumolysin, that is, hemolytic activity, etc.

Inactive Publication Date: 2007-01-24
EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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Problems solved by technology

The advantage is that it has a large number of valences and a wide range of protection, but studies have shown that this type of vaccine has the following problems: (1) Capsular polysaccharide is a T cell-independent antigen, although it can induce protective IgM antibodies after the initial vaccination, the effect is not effective. The price is not high, and re-vaccination cannot induce the required T cell memory and high-efficiency IgG antibodies; (2) The immune response produced is only effective against adult infection, but less effective against pneumococcal otitis media in children under 2 years old
However, th

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  • Pneumococcus polysaccharide protein coupling vaccine and its preparing method
  • Pneumococcus polysaccharide protein coupling vaccine and its preparing method
  • Pneumococcus polysaccharide protein coupling vaccine and its preparing method

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Embodiment Construction

[0021] The (pneumococcal) hemolysin protein produced by pneumococcus itself is very small. The present invention uses genetic engineering methods to clone and obtain the pneumolysin gene from Streptococcus pneumoniae. After modification, the pneumolysin protein is expressed in E. It is purified.

[0022] 1. The present inventor has initially designed a pneumolysin gene sequence (see SEQ ID NO.3 in the sequence listing) (GenBank accession No.X52474) according to the report of Walker et al. in 1987. CATATG ) and XhoI (C TCGAG ) of the primer sequence 4 (SEQ ID NO.4) and primer sequence 5 (SEQ ID NO.5) of the recognition site, using PCR technology to amplify from the chromosomal DNA of pneumococcus to obtain pneumococcal hemolytic bacteria with the sequence of SEQ ID NO.3 pneumolysin gene, and the pneumolysin gene and the prokaryotic expression vector (plasmid) pET-21a were digested with restriction endonucleases NdeI and XhoI respectively, and a gene recombinant plasmid capab...

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Abstract

The present invention is pneumococus polysaccharide protein coupling vaccine comprising covalently connected pneumococus capsule polysaccharide and recombinant pneumolysin without hemolytic activity modification and its preparation process. The vaccine has pneumolysin without hemolytic activity as protein carrier, no need of eliminating hemolysis toxicity of pneumolysin with formalin and ensured safety, and may be used for infant below 2 yeas old to prevent tympanitis. Owing to the pneumolysin as the self protein, the vaccine has no probable immune interference reaction and strengthened immune protecting effect. The vaccine has cross immunizing protection effect on various kinds of serum type pneumococus and raised immune memory response to pneumococus infection.

Description

technical field [0001] The present invention relates to the category of novel vaccines in the field of biomedicine, in particular to the development of pneumococcal polysaccharide-protein conjugated vaccines using genetic engineering technology, and more specifically to pneumococcal polysaccharides with modified pneumolysin (protein) without hemolytic activity as the carrier Protein conjugate vaccine, its preparation method and application as otitis media vaccine. Background technique [0002] Otitis media (including acute otitis media (AOM) and secretory otitis media (OME)) is one of the most common diseases in infants and young children. It is an inflammatory disease of the middle ear caused by bacteria or viruses. Studies have found that more than 80% of children aged 2-3 have suffered from otitis media at least once, and about 17% of children have had 3 or more episodes of otitis media from birth to 6 months of age. From birth to 1 year old, 62% of children suffer from ...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P27/16
Inventor 王正敏陈兵
Owner EYE & ENT HOSPITAL SHANGHAI MEDICAL SCHOOL FUDAN UNIV
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