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Preparation method and kit of dendritic cell vaccine loaded with tumor-specific antigen epitope polypeptide

A tumor-specific antigen, dendritic cell technology, applied in anti-tumor drugs, medical preparations containing active ingredients, pharmaceutical formulations, etc., can solve the problems of cancer recurrence, poor prognosis, and inability to obtain patients, and achieve a radical cure for recurrence. and transfer, the effect of overcoming tolerance

Inactive Publication Date: 2017-03-22
李金珍
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology, that is, the conventional treatment methods such as surgery, chemotherapy and radiotherapy can remove or kill most of the malignant tumors in the body, but there are still some residual tumor cells or tumor cells that develop drug resistance and become the "culprit" of cancer recurrence and metastasis, making Patients can not get a long survival time and poor prognosis

Method used

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  • Preparation method and kit of dendritic cell vaccine loaded with tumor-specific antigen epitope polypeptide
  • Preparation method and kit of dendritic cell vaccine loaded with tumor-specific antigen epitope polypeptide
  • Preparation method and kit of dendritic cell vaccine loaded with tumor-specific antigen epitope polypeptide

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0045] Preparation Example 1 Prediction of Human Epithelial Cell Adhesion Molecule HLA-A2010 Positive Epitope Polypeptide

[0046] Using the Bioinformatics and Molecular Analysis System (BIMAS) to predict the binding of HLA peptides, screen the HLA epitope peptides that specifically bind to human epithelial cell adhesion molecules, and select 6 amino acid sequences that have a binding affinity of more than 100 to HLA-A0201 , see Table 1:

[0047] ranking start site Peptide Amino Acid Sequence binding affinity score 1 174 YQLDPKFIT 125.377 2 184 ILYEN NVIT 68.146 3 260 SMQGLKAGV 50.232 4 11 LLLAAATAT 46.873 5 271 VIVVVVIAV 37.393 6 5 QVLAFGLLL 26.281

preparation example 2

[0048] Preparation Example 2 Dendritic Cell Acquisition

[0049] Collect 2-4 liters of peripheral blood from tumor patients with a blood cell separator and purify the mononuclear cells with the lymphocyte separation medium; after counting by the cell counter, the mononuclear cells are diluted to 3-5×10 with AIM-V medium 6 , add 75cm 2 Attached to the wall in the culture bottle;

[0050] At 37°C, 5% CO 2 After incubation in the incubator for 90 minutes, non-adherent cells were washed and collected for activating T lymphocytes; adherent cells were induced by adding complete AIM-V medium, containing 5% autologous serum, 1000IU / mL recombinant human granules Macrophage colony-stimulating factor and 500IU / mL recombinant human interferon-α;

[0051] Harvest immature dendritic cells (DCs) on the fourth day of culture; mature DCs were induced to the fifth day with complete AIM-V medium, containing 5% autologous serum, 1000IU / mL recombinant human granulocyte macrophage colonies Stim...

Embodiment 3

[0052] Example 3 EpCAM epitope polypeptide-loaded DCs

[0053] Cultured to the fourth day, 3×10 6 An immature DCs with epithelial cell adhesion molecule protein SEQ ID NO.1-6 more than one epitope polypeptide composition at 37 ° C, 5% CO 2 Load in the incubator for 2-4 hours; induce culture with full AIM-V medium until the fifth day, and harvest the mature antigen polypeptide-loaded DCs by centrifugation (10min, 1000rpm); the obtained DCs are washed 3 times with normal saline, and then resuspended into normal saline to a concentration of 5×10 6 mature DC / mL, and add human albumin with a final concentration of 1% by mass to volume to prepare a DC vaccine loaded with the EpCAM antigen epitope polypeptide composition.

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Abstract

The invention provides a preparation method for a dendritic cell vaccine loaded by a tumor specific antigenic epitope polypeptide. The method comprises the following steps: adopting a human leucocyte antigen A201 positive epitope of an epithelial cell adhesion molecule identified by tumor specific expression, namely a polypeptide composed of amino acid sequences represented by any one of SEQ ID NO.1-6; loading dendritic cells by using the polypeptide of any one of the SEQ ID NO.1-6 to prepare the dendritic cell vaccine. The dendritic cell vaccine can be used for initiating a very strong target anti-tumor cytotoxic T lymphocyte effect. The invention further provides a kit for preparing the dendritic cell vaccine loaded by the tumor specific antigenic epitope polypeptide.

Description

technical field [0001] The invention relates to a method for preparing a dendritic cell vaccine loaded with a tumor-specific antigen epitope polypeptide, a kit using the method, and a dendritic cell and vaccine obtained by the method. In particular, the present invention relates to a human leukocyte antigen A201-positive epitope polypeptide using the identified tumor-specifically expressed epithelial cell adhesion molecule, a preparation method for loading dendritic cells, a kit using the method, and a method for using the method Obtained dendritic cell vaccine. Background technique [0002] At present, malignant tumor is one of the most important diseases threatening human health. There are 22 million cancer patients in the world, and 10 million new cases are added every year. There are 2.8 million new cancer cases in China every year, and the number of cancer cases is increasing at an average annual rate of 3%-5%. The existing technology, that is, the conventional treatm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61K47/42A61K45/00A61P35/00A61P35/04
Inventor 金光瑞王琳
Owner 李金珍
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