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Preparation method of tumor dendritic cell vaccine sensitized by glycosylated MUC1 (mucoprotein 1) antigen

A technology for vaccine preparation and glycosylation, which is applied in the field of preparation of anti-tumor immune cell vaccines, and can solve the problem of low efficiency of antigen presentation.

Inactive Publication Date: 2013-10-30
SHANGHAI FUHETAI BIOLOGICAL TECH CO LTD
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Problems solved by technology

However, this method has low efficiency of DC cells taking up antigens and presenting antigens, and it is easy to cause side effects such as immune tolerance

Method used

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  • Preparation method of tumor dendritic cell vaccine sensitized by glycosylated MUC1 (mucoprotein 1) antigen
  • Preparation method of tumor dendritic cell vaccine sensitized by glycosylated MUC1 (mucoprotein 1) antigen

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Embodiment Construction

[0022] The specific technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments, so that those skilled in the art can understand the present invention without restricting their rights.

[0023] 1. Preparation of MUC1M-FP (such as figure 1 Shown):

[0024] 1) Preparation of MUC1FP: A 309 base fragment (pDF9.3, encoding a 60 base motif fragment in the VNTR region of MUC1 cDNA slightly more than 5) was cloned into the bacterial expression vector pGEX-3X for correct expression Frame and express direction. MUC1-FP (including GST and MUC1VNTR) was induced with IPTG. The cells were collected by centrifugation, washed and sonicated (in a buffer containing TritonX-100), the supernatant containing the solubilized protein MUC1-FP was mixed with glutathione-coated magnetic beads (disulfide bond connection), and then collected by centrifugation . The fusion protein MUC1-FP was washed with a buffer containing reduc...

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Abstract

The invention provides a preparation method of a tumor dendritic cell vaccine sensitized by glycosylated antigen MUC1 (mucoprotein 1). Glycosylation modified tumor antigen peptide is used for sensitizing externally cultured DC (Dendritic Cell) and then the activated DC cells are transfused to a tumor patient. The invention further provides an application of the tumor DC cell vaccine sensitized by the glycosylated MUC1 antigen in treatment of breast cancer and pancreatic cancer. The preparation method provided by the invention has the following advantages (1) the DC cell vaccine amplification efficiency is high; (2) the DC cells have high antigen activity; (3) the DC cells have high maturity degree, and side effects such as immune tolerance are prevented; and (4) small side effects are caused.

Description

Technical field [0001] The invention relates to a method for preparing an anti-tumor immune cell vaccine, in particular to a method for preparing an anti-tumor immune dendritic cell vaccine. Background technique [0002] Dendritic cells (DC) are the most powerful antigen-presenting cells in the human body. They can ingest and process antigens, present antigens in the form of MHC I and II peptide conjugates, and promote the proliferation of T and B lymphocytes. The ability to play an extremely important role in both innate immunity and acquired immunity. [0003] Early animal experiments and clinical trials have shown that DC vaccines loaded with appropriate forms of tumor antigens, whether proteins, peptides, or nucleic acids, can activate antigen-specific T cells that can recognize and kill tumor cells, and can produce immune memory effects. . At present, there are three main types of DC vaccines: DCs modified with cellular tumor antigens; DCs modified with tumor antigen peptide...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00C12N5/0784
Inventor 方存磊
Owner SHANGHAI FUHETAI BIOLOGICAL TECH CO LTD
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