Recombination antigen composition, vaccine and carrier and method for preparing antigen composition

A composition and antigen technology, applied in the field of immunization, can solve the problems such as the huge effect of antigen vaccines, the difficulty in producing protective effects, and the difficulty in producing immune responses for vaccines.

Inactive Publication Date: 2013-06-05
SHANGHAI UNITED CELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the process of vaccine research and development, people found that the choice of antigen has a great influence on the effect of the vaccine, and it is oft

Method used

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  • Recombination antigen composition, vaccine and carrier and method for preparing antigen composition
  • Recombination antigen composition, vaccine and carrier and method for preparing antigen composition
  • Recombination antigen composition, vaccine and carrier and method for preparing antigen composition

Examples

Experimental program
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preparation example Construction

[0029] The preparation of candidate vaccines requires effective antigens, and the use of effective antigens to immunize the body can stimulate the body to generate a systemic immune response, and then prevent the body from infection.

[0030] In the preparation of the antigen used in the vaccine, the present invention adopts the CT-A2-CT-B system to increase the volume of the antigen and utilizes the immune adjuvant function of CT to enhance the immunogenicity of the antigen. Antigens that can use this system include, but are not limited to: Helicobacter pylori-related antigens, typhoid antigens, influenza HA antigens, and the like.

[0031] For these antigens, fusion proteins of corresponding antigens and CT-A2 can be prepared by methods known in the art, and then used together with CT-B protein to achieve the purpose of enhancing immunogenicity.

[0032] Taking the preparation of Helicobacter pylori-related vaccines as an example, the inventors preferentially selected Helico...

Embodiment 1

[0038] The preparation of embodiment 1UreB-CTA2 / 5CTB

[0039] 1.1 Construction of recombinant engineering bacteria PET-28a-UreB-CTA2 / BL21-DE3

[0040] 1.1.1 Construction of fusion gene UreB-CTA2:

[0041] The amino acid sequence of UreB comes from Helicobacter pylori strain MEL-HP27, the amino acid sequence of CTA2 comes from Vibrio cholerae O395 strain, and the UreB-CTA2 gene is codon-optimized and synthesized.

[0042] The UreB-CTA2 gene sequence (SEQ ID NO.: 1) is:

[0043] ATGAAAAAAAATCTCTAGGAAAGAGTACGTAAGCATGTACGGTCCGACCAC 50

[0044] CGGTGACAAAGTTCGTCTGGGTGACACCGACCTGATCGCTGAAGTTGAAC 100

[0045]ACGACTACACCATCTACGGTGAAGAACTGAAATTCGGTGGTGGTAAAACC 150

[0046] CTGCGTGAAGGTATGTCTCAGTCTAACAACCCGTCTAAAGAAGAACTGGA 200

[0047] CCTGATCATCACCAACGCTCTGATCGTTGACTACACCGGTATCTACAAAAG 250

[0048] CTGACATCGGTATCAAAGACGGTAAAATCGCTGGTATCGGTAAAAGGTGGT 300

[0049] AACAAAGACATGCAGGACGGTGTTAAAAACAACCTGTCTGTTGGTCCGGC 350

[0050] TACCGAAGCTCTGGCTGGTGAAGGTCTGATCGTTACCGCTGGTGGTATCG 4...

Embodiment 2

[0110] The preparation method of embodiment 2CagA-CTA2 / 5CTB

[0111] CagA-CTA2 / 5CTB was prepared by the same steps as in Example 1, except that the CagA gene was used instead of the UreB gene.

[0112] The corresponding gene sequence (SEQ ID NO.: 2) is:

[0113] ATGAAACTGTTCGGTAACTCTAACAACAACAACAACGGTCTGAAAAACGA 50

[0114] ACCGATCTACGCTCAGGTTAACAAAAAAAAAGCTGGTCAGGCTACCTCTC 100

[0115] CGGAAGAACCGATCTACGCTCAGGTTGCTAAAAAAGTTCTGCTAAAATC 150

[0116] GACCAGCTGAACGAAGCTACCTCTGCTATCAACCGTAAAATCGACCGTAT 200

[0117] CAACAAAATCGCTTCTGCTGGTAAAGGTGTTGGTGGTTTCTCTGGTGCTG 250

[0118] GTCGTTCTGCTTCTCCGGAACCGATCTACGCTACCATCGACTTCGACGAA 300

[0119] GCTAACCAGGCTGGTTTCCCGCTGCGTCGTTCTACCGCTGTTAACGACCT 350

[0120] GTCTAAAGTTGGTCTGTCTCGTGAACAGGAACTGACCCGTCGTATCGGTG 400

[0121] ACCTCAAACCAGGCTGTAAGCGAAGCTAAAACCGGTCACTTCGACAAACTG 450

[0122] GAACAGAAAATGGACGAACTGAAAGACTCTACCAAAAAAAACGCTCTGAA 500

[0123] ACTGTGGGCTGAATCTACCAAACAGGTTCCGACCGGTCTGCAGGCTAAAC 550

[0124] TGGACAACTACGCTA...

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Abstract

The invention provides a recombination antigen composition, a vaccine and a carrier and a method for preparing the antigen composition. A recombination helicobacter pylori antigen which comprises a helicobacter pylori antigen, the fusion proteins of a cholera toxin CT-A2 subunit and cholera toxin CT-B proteins is optimized. The compatibility of the CT-A2 and the CT-B proteins is used, a chimeric structure of CT-A2-5CT-B is formed, and accordingly an antigen with higher titer is formed through the helicobacter pylori antigen and the fusion proteins of the cholera toxin CT-A2 subunit. Meanwhile, the effect that the CT-B enhances immunization is used, and the effect that the antigen immunogenicity of the recombination antigen composition is enhanced is achieved. In addition, the chimeric protein constituted by the recombination antigen stimulates a mucous membrane to generate secreting type IgA immunization.

Description

technical field [0001] The invention relates to the field of immunization, especially the field of recombinant antigen, especially the antigen, vaccine, preparation method and carrier for Helicobacter pylori infection immunity. Background technique [0002] In the field of vaccine preparation, many kinds of antigens cannot stimulate sufficient immune response due to reasons such as low self-antigenicity or too small molecules, thus limiting the possibility of vaccine development and the effectiveness of vaccines. In order to overcome this difficulty, many methods are currently used to increase the immunogenicity of the antigen, for example, the use of immune adjuvants. Commonly used adjuvants can be divided into four categories: inorganic adjuvants, such as aluminum hydroxide, alum, etc.; organic adjuvants, microorganisms and their products such as mycobacteria (Mycobacterium tuberculosis, BCG), Brevibacterium, pertussis bacillus, endotoxin, Bacterial extracts (muramoyl dip...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K39/02A61K39/112A61K39/145A61K39/39A61P31/04A61P31/16
CPCY02A50/30
Inventor 耿雨红郭鸿郑云程马唯虹
Owner SHANGHAI UNITED CELL BIOTECH
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