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GM-CSF-HER2 recombinant protein, and preparation method and applications thereof

A technology of GM-CSF-HER2 and recombinant protein, which is applied in the field of GM-CSF-HER2 recombinant protein and its application, can solve the problems of premature apoptosis and low antigen specificity of DC cells, and achieve the improvement of antigen specificity, Effect of enhanced specificity and increased lifespan

Inactive Publication Date: 2014-04-30
SHANGHAI CLAISON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the shortcomings of DC cells induced by conventional methods at this stage, such as low antigen specificity and premature apoptosis

Method used

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  • GM-CSF-HER2 recombinant protein, and preparation method and applications thereof
  • GM-CSF-HER2 recombinant protein, and preparation method and applications thereof
  • GM-CSF-HER2 recombinant protein, and preparation method and applications thereof

Examples

Experimental program
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Embodiment 1

[0045] The specific coding sequence of human breast cancer cell SKBr3 specific antigen HER2 was recombined with the coding sequence of granulocyte colony-stimulating growth factor GM-CSF.

[0046] Among them, the granulocyte colony-stimulating growth factor GM-CSF is from NCBI, its sequence number is AAA52578, and its coding sequence is:

[0047] tqpwehvnai qearrllnls rdtaaemnet vevisemfdl qeptclqtrl elykqglrgs 60

[0048] ltklkgpltm mashykqhcp ptpetscatq iitfesfken lkdfllvipf dc 112

[0049] Add restriction sites HindⅢ and Not I to both ends of the GM-CSF gene sequence;

[0050] Human breast cancer cell SKBr3-specific antigen HER2 is from NCBI, its sequence number is NP_004439, and its coding sequence is:

[0051] gcpaeqrasp ltsiisavvg illvvvlgvv fgilikrrqq kirk 44

[0052] Add restriction sites Xho I and BamH I at both ends of the HER2 gene sequence;

[0053] Using gene recombination technology, the above two gene fragments were cloned into plasmid pCEP4 to construct a r...

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Abstract

The invention relates to a GM-CSF-HER2 recombinant protein, and a preparation method and applications thereof, and specifically relates to a GM-CSF-HER2 recombinant protein which can be used for preparation of antitumor drugs. Amino acid sequence of the GM-CSF-HER2 recombinant protein is represented by SEQ ID No.1. The preparation method comprises following steps: gene sequences of GM-CSF and HER2 are cloned into plasmid pCEP4 via enzyme digestion for recombinant plasmid construction; human embryo kidney 293 cells are transfected by the obtained recombinant plasmid, and recombinant protein expression is performed; and the obtained recombinant protein is purified and collected. The recombinant protein can be used for producing drugs used for treating cancers such as breast cancer. According to the preparation method, genetic engineering method is adopted; tumor specific antigen, transducin, and GM-CSF which is a key agent for in vitro induction of DC cells are combined; the fusion protein which possesses specificity and is capable of prolonging DC cell life span is prepared; prolonging of DC cell survival time and starting of specific immune response are ensured; the GM-CSF-HER2 recombinant protein is capable of avoiding additional adding of GM-CSF in routine DC cell culturing, and insufficient of GM-CSF in culturing cycles; and coexpression GM-CSF is capable of promoting cytotoxic effects.

Description

[technical field] [0001] The present invention relates to a GM-CSF-HER2 recombinant protein and its method and application, in particular to a GM-CSF-HER2 recombinant protein capable of being used as a drug for treating tumors, its preparation method, and its DC vaccine preparation method and application. [Background technique] [0002] One of the fundamental reasons for the occurrence and development of tumors is that the body's immune system cannot effectively control its growth. To correct this, so that the body's immune system can effectively generate immune rejection against tumor cells has been studied for nearly a hundred years. topic. Tumor vaccines are the main content of research in this area. Among many tumor vaccines, dendritic cells (dendritic cells, DC) tumor vaccines designed according to the important role of antigen-presenting cells in tumor immunity are currently the most popular research direction. . The body's immune response is first captured, processe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/85A61K39/00A61P35/00C12N5/0784
Inventor 张超叶永清苏国新钱妮
Owner SHANGHAI CLAISON BIOTECH
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