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Preparation method for tumor antigen targeting DC cells

A tumor antigen, targeted technology, applied in the field of cell culture, to achieve the effect of strong tumor-specific immune response, wide variety, and efficient uptake

Inactive Publication Date: 2016-04-13
英普乐孚生物技术(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to improve the ability of DC cells to absorb antigens, and solve the problem of how to allow DC cells to specifically absorb specific tumor-specific antigens when the concentration of tumor-specific antigens is very low or there are many other non-tumor-specific antigens Even unknown tumor-specific antigens, thereby improving the specificity and efficacy of DC cells in treating tumors

Method used

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  • Preparation method for tumor antigen targeting DC cells
  • Preparation method for tumor antigen targeting DC cells
  • Preparation method for tumor antigen targeting DC cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Step 1: Prepare Tumor Tissue Lysate

[0040] 1.1 Take fresh in situ resected gastric cancer tissue positive for CEA.

[0041] 1.2 Remove the normal tissue above the surgically resected tumor tissue, rinse with normal saline until there is no blood and no fat residue.

[0042] 1.3 Soak with 0.1% chlorhexidine (chlorhexidine) for 30 minutes, rinse with sterile saline 3 to 4 times.

[0043] 1.4 Shred the tumor tissue with sterile surgical scissors, add an appropriate amount of saline for grinding, and collect the single cell suspension after filtering through a 200-mesh mesh.

[0044] 1.5 Put the single-cell suspension into a centrifuge tube, freeze at -86°C, thaw at room temperature, repeat freezing and thawing 10 times, and observe under a microscope until no cells survive.

[0045] 1.6 Centrifuge the single cell lysate at 3000rpm for 30min. Aseptic operation The supernatant was filtered and sampled with a 0.22 μm pore size sterile filter and stored at 4°C for quality...

Embodiment 2、3

[0056] The operation process of steps 1-5 is the same as that of Example 1, except that the concentration and time of adding CEA antibody, tumor tissue lysate in step 3 are different:

Embodiment 2

[0057] Example 2: DC cells were cultured until the nth day (n=3), the concentration of anti-CEA antibody was 20 ng / ml, and the concentration of tumor tissue lysate was 100 ug / ml.

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Abstract

The invention relates to a preparation method for tumor antigen targeting DC cells. By means of the characteristic that an antibody can be combined with a tumor antigen and a DC cell surface FC receptor, the DC cells can specifically take in tumor cell debris through the antibody, so that the tumor cell specific antigen can be taken in and presented. Compared with a method for directly incubating DC cells and tumor cell lysates together, the preparation method for the tumor antigen targeting DC cells has the specificity for taking in the tumor cell antigen and is higher in efficiency, and therefore more and stronger tumor cytotoxicity lymphocytes.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and relates to a method for separating and culturing tumor antigen-targeted DC cells. Background technique [0002] DC cells are professional antigen-presenting cells (Antigenpresenting cells, APCs) with the strongest function in the body, which can efficiently absorb, process and present antigens. DC cells present antigens to T cells for recognition through MHC molecules. Once the antigens presented by MHC molecules are recognized, activation and expansion of this type of T cells will occur. After activation, CD8+ T cells will kill target cells expressing this antigen. Therefore, using this characteristic of DC cells, DC cells can be loaded with tumor cell-specific antigens, and presented by DC cells to T cells, thereby activating tumor-specific T cells, which can kill tumor cells. [0003] Therefore, how to make DC cells obtain tumor cell-specific antigens is the key to this technology. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/0783A61K35/15A61P35/00
CPCC12N5/0639A61K35/15C12N5/0636
Inventor 谢志明武宁郑蓉蓉熊青卉杨春辉
Owner 英普乐孚生物技术(上海)有限公司
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