Anti-tumor chimeric protein, anti-tumor vaccine, and anti-tumor vaccine for nasal mucosa administration

A chimeric protein, anti-tumor technology, applied in the field of anti-tumor vaccines, can solve the problems of weak immunogenicity, inability to perform immunotherapy, inability to recognize and present to activate toxic T lymphocytes, etc.

Inactive Publication Date: 2019-01-15
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a major problem in the development of tumor vaccines is that the immunogenicity of tumor antigens is weak, and they canno

Method used

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  • Anti-tumor chimeric protein, anti-tumor vaccine, and anti-tumor vaccine for nasal mucosa administration
  • Anti-tumor chimeric protein, anti-tumor vaccine, and anti-tumor vaccine for nasal mucosa administration
  • Anti-tumor chimeric protein, anti-tumor vaccine, and anti-tumor vaccine for nasal mucosa administration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] see figure 1 , this example provides a recombinant engineered bacterium containing the first polypeptide sequence, marked as pET22b-GM-CSF-CTA2-TAT, the construction method is as follows:

[0050] In this example, the commercial carrier pET22b was selected, and the sequence of GM-CSF-CTA2-TAT (GM-CSF, CTA2 subunit sequence of cholera toxin and TAT sequence of cell penetrating peptide sequence) was artificially synthesized and cut with NdeI and XhoI Insert it into the pET22b plasmid to obtain the recombinant plasmid pET22b-GM-CSF-CTA2-TAT for fusion expression, transform Escherichia coli BL21(DE3), and obtain the recombinant engineering bacteria containing the first polypeptide sequence.

[0051] Wherein, the amino acid sequence of GM-CSF-CTA2-TAT is:

[0052] MAPTRSPITVTRPWKHVEAIKEALNLLDDMPVTLNEEVEVVSNEFSFKKLTCVQTRLKIFEQGLRGNFTKLKGALNMTASYYQTYCPPTPETDCETQVTTYADFIDSLKTFLTDIPFECKKPSQKDPTGGASEFELGGGGSMSNTCDEKTQSLGVKFLDEYQSKVKRQIFSHRPHPDLEGTHNRIKDERLVD. The connection bet...

Embodiment 2

[0060] see figure 1 , this embodiment provides a recombinant engineered bacterium containing the second polypeptide sequence, marked as pET28a-CTB-PSMA624-632, the construction method is as follows:

[0061] In this example, the commercial vector pET28a was selected, and the amino acid sequence of CTB was recombined with pET28a through transformation to obtain the recombinant plasmid pET28a-CTB. The PSMA624-632 gene was cut with NheI and XhoI and inserted into the downstream of the CTB gene to form a recombinant plasmid for fusion expression. pET28a-CTB-PSMA624-632 was transformed into Escherichia coli BL21(DE3) to obtain a recombinant engineered bacterium containing the second polypeptide sequence.

[0062] The second polypeptide is a fusion protein, and the amino acid sequence denoted as (CTB-PSMA624-632)5 is:

[0063] MTPQNITDLCAEYHNTQIYTLNDKIFSYTESLAGKREMAIITFKNGAIFQVEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMANASGGTYSVSFDSLLEHHHHHH. SEQ ID No: 1 is connected to ...

Embodiment 3

[0070] This embodiment provides the expression method of the fusion protein GM-CSF-CTA2-TAT:

[0071] Cultivate the recombinant engineered bacteria containing the first polypeptide sequence of Example 1 at 37°C, with ampicillin and 0.1mmol / L isopropyl-β-D-thiopyran galactobacter at a final concentration of 50 μg / mL Glycoside (IPTG) induces the expression of the fusion protein to obtain the first polypeptide (GM-CSF-CTA2-TAT).

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Abstract

The invention particularly relates to an antitumor chimeric protein, an antitumor vaccine and an antitumor vaccine for nasal mucosa administration, belonging to the technical field of biomedicine. Theanti-tumor chimeric protein provided by the invention comprises: a first polypeptide, which comprises a lymphokine sequence for enhancing an immunoreaction and a CTA2 subunit sequence of cholera toxin; and a second polypeptide, which comprises a CTB subunit sequence of the cholera toxin and a tumor-specific epitope peptide sequence, wherein a CTA2 subunit of the cholera toxin in the first polypeptide and a CTB subunit of the cholera toxin in the second polypeptide are mutually chimeric to form the anti-tumor chimeric protein. The anti-tumor chimeric protein, the anti-tumor vaccine and the anti-tumor vaccine for nasal mucosa administration provided by the invention are used for solving the technical defect of poor immunogenicity of tumor vaccines in the prior art.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an anti-tumor chimeric protein, an anti-tumor vaccine and an anti-tumor vaccine for nasal mucosa administration. Background technique [0002] Tumor vaccines are a hot spot in oncology research in recent years. Through the administration of tumor-specific antigens or antigenic polypeptides, they can activate, restore or strengthen the body's anti-tumor immune response, and then kill and eliminate tumor cells expressing tumor-specific antigens. However, a major problem in the development of tumor vaccines is that the immunogenicity of tumor antigens is weak, and they cannot be recognized and presented by the immune system to activate toxic T lymphocytes (CTL), and cannot play the role of immunotherapy. Contents of the invention [0003] The purpose of the present invention is to provide a safe and immunogenic anti-tumor chimeric protein, an anti-tumor vaccine and...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/00A61K39/39A61P35/00
CPCA61K39/00A61K2039/543A61K2039/55522A61K2039/55544A61P35/00C07K14/28C07K14/4748C07K14/535C07K2319/10C07K2319/40C07K2319/55
Inventor 王华倩林丹敏何华锋宋彩燕孙家杰龙威郑希杜志云赵肃清张琨
Owner GUANGDONG UNIV OF TECH
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