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Polysaccharide conjugate vaccine and preparation method thereof

A technology combining vaccines and polysaccharides, applied in the field of vaccines, can solve problems such as bacterial polysaccharide conjugates that have not yet been seen

Active Publication Date: 2016-03-30
BEIJING BIOLOGICAL PROD INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacterial polysaccharide conjugates using such proteins as carrier proteins have not been seen so far

Method used

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  • Polysaccharide conjugate vaccine and preparation method thereof
  • Polysaccharide conjugate vaccine and preparation method thereof
  • Polysaccharide conjugate vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The present invention will be described in detail below by taking the combination of Escherichia coli heat-labile enterotoxin B subunit (LTB) and group C meningococcal polysaccharide (GCMP) as an example.

[0037] (1) Cloning and expression of recombinant Escherichia coli heat-labile enterotoxin B subunit (rLTB) 1) Synthesis of LTB gene and construction of expression vector

[0038] The LTB gene sequence is selected from GENBANK, and the sequence number is GI: 157021177. According to the Escherichia coli codon usage frequency table (references: Maloy, S., V.Stewart, and R.Taylor.1996.Geneticanalysisofpathogenicbacteria.ColdSpringHarborLaboratoryPress, NY.) Optimize this sequence, add 6 histidine tags at the C-terminus of the sequence and stop codon, and then add NdeI and AvrII enzyme cutting sites at both ends of the sequence respectively.

[0039] Artificially synthesize the above-mentioned LTB gene, then use NdeI and AvrII to double-enzyme digest LTB and pET42b (purc...

Embodiment 2

[0090] The present invention will be described in detail below by taking the combination of Campylobacter jejuni flagellar secretory protein A1 (FspA1) and group A meningococcus polysaccharide (GAMP) as an example.

[0091] (1), Cloning and expression of FspA1

[0092] 1) Synthesis of FspA1 gene and construction of expression vector

[0093] The FspA1 gene sequence is selected from GENBANK, and the sequence number is GI: 116292639. According to the Escherichia coli codon usage frequency table (references: Maloy, S., V.Stewart, and R.Taylor.1996.Geneticanalysisofpathogenicbacteria.ColdSpringHarborLaboratoryPress, NY.) Optimize this sequence, add 6 histidine tags at the C-terminus of the sequence and stop codon, and then add NdeI and SacI restriction sites at both ends of the sequence.

[0094] The above-mentioned FspA1 gene was artificially synthesized, and then FspA1 and pET30a (purchased from Novagen) were double-digested with NdeI and SacI, and then the target fragment aft...

Embodiment 3

[0122] The present invention will be described in detail below by taking the combination of pneumococcal surface membrane protein A (PspA) and group A meningococcal polysaccharide (GAMP) as an example.

[0123] (1), cloning and expression of recombinant PspA

[0124] 1) Synthesis of PspA gene and construction of expression vector

[0125] The PspA gene sequence is selected from GENBANK, and the sequence number is GI:193804931. According to the Escherichia coli codon usage frequency table (references: Maloy, S., V.Stewart, and R.Taylor.1996.Geneticanalysisofpathogenicbacteria.ColdSpringHarborLaboratoryPress, NY.) Optimize this sequence, add 6 histidine tags at the C-terminus of the sequence and stop codon, and then add NdeI and SacI restriction sites at both ends of the sequence.

[0126] The above-mentioned PspA gene was artificially synthesized, and then PspA and pET30a (purchased from Novagen) were double-digested with NdeI and SacI, and then the target fragment after doub...

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Abstract

The invention relates to a polysaccharide conjugate vaccine, which comprises a chemical conjugate consisting of a bacterial polysaccharide and a vector protein, wherein the vector protein is a bacterial slime invasion-associated protein, such as a bacillus coli heat-labile toxin B subunit, a campylobacter jejuni flagellum secretion protein A1, a campylobacter jejuni flagellum secretion protein A2, a pneumococcus surface protein and a pneumococcus surface adhesin. The invention further relates to a preparation method for the polysaccharide conjugate vaccine. According to the invention, the mucosa delivery of the polysaccharide conjugate vaccine can be realized on the premise of not adding any other extrinsic protein; and due to the adoption of an immune route, the vaccine is more efficient, convenient and safe to use.

Description

technical field [0001] The invention relates to the field of vaccines, more specifically, the invention relates to polysaccharide conjugated vaccines and a preparation method thereof. Background technique [0002] The capsular polysaccharide of bacteria is an important protective antigen. It is one of the important measures in the history of vaccine development to use chemically purified bacterial capsular polysaccharide component vaccines to prevent infectious diseases. Among them, the polysaccharide vaccines that have been widely used all over the world include epidemic meningococcal A, C, 4-valent polysaccharide vaccines such as Y and W135, and pneumococcal 23-valent polysaccharide vaccines. However, polysaccharide vaccines have great limitations. They have poor immunogenicity in infants under 2 years old and cannot induce effective immune protection. A vaccine prepared chemically with bacterial polysaccharide (PS) and a protein carrier is called a conjugate vaccine. A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/09A61K39/145A61K47/48A61P31/04A61P31/12
CPCY02A50/30
Inventor 李启明张学峰马智静唐玉龙张靖陈实靳玉琴卫江波
Owner BEIJING BIOLOGICAL PROD INST CO LTD
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