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Specific targeting receptor protein MAGEA and construction method of CAR-T carrier thereof

A technology of receptor protein and construction method, which is applied in the direction of tumor-specific antigen, mammalian protein, chemical instruments and methods, etc., can solve the problem of lack of effective targets, etc., and achieve the effect of long-lasting immunity

Inactive Publication Date: 2017-12-08
赵蔚
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Problems solved by technology

The ideal state of CAR-T cell therapy is that T cells can clearly distinguish the target molecules of tumor cells and normal tissues. Unfortunately, CAR-T cells currently lack effective targets for the treatment of solid tumors, especially in the immunotherapy of osteosarcoma.

Method used

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Embodiment Construction

[0015] In order to facilitate those skilled in the art to understand, the present invention will be described in further detail below in conjunction with embodiment:

[0016] A method for constructing a specifically targeting receptor protein MAGEA and its CAR-T carrier, the construction method comprising first extracting the RNA of monoclonal hybridoma cells against osteosarcoma, and then using specific primers for monoclonal antibodies to perform RT-PCR, cloning the variable region DNA fragments of the heavy and light chains of the antibody into the T vector, sequencing to obtain the receptor protein MAGEA that specifically recognizes tumor cells, and cloning its expression sequence into the CAR-T vector, A novel CAR-T vector specifically targeting MAGEA was obtained.

[0017] The nucleotide sequence NCBI code of the receptor protein MAGEA that specifically recognizes tumor cells is NM_005362.3.

[0018] The NCBI code of the amino acid sequence of the receptor protein MAGEA...

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Abstract

The present invention provides a construction method for specifically targeting the receptor protein MAGEA and its CAR-T carrier. The construction method includes first extracting the RNA of monoclonal hybridoma cells against osteosarcoma, and then using the specific monoclonal antibody RT-PCR with specific primers, clone the variable region DNA fragments of the heavy and light chains of the antibody into the T vector, sequence, obtain the receptor protein MAGEA that specifically recognizes tumor cells, and clone its expression sequence into CAR-T On the carrier, a new type of CAR‑T carrier specifically targeting MAGEA was obtained. The present invention can provide an effective target for solid tumors treated by CAR-T cells, especially in the immunotherapy of osteosarcoma.

Description

technical field [0001] The invention relates to the field of tumor immunotherapy, in particular to a method for constructing a specific targeting receptor protein MAGEA and a CAR-T carrier thereof. Background technique [0002] Malignant tumors are currently one of the leading causes of death worldwide. "Global Cancer Report 2014" stated that in 2012, the number of cancer patients and deaths worldwide was disturbingly increasing. Nearly half of the new cancer cases occurred in Asia, most of which were in China, and China ranked first in the number of new cancer cases. . Among the four types of malignant tumors including liver, esophagus, stomach and lung, China ranks first in the world in terms of new cases and deaths. The treatment process of cancer is quite complicated, and all current treatment methods have certain limitations, and cellular immunotherapy has become the only possible method to completely eliminate cancer cells in theory: the immune response can find an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/64C12N15/12C07K14/705C07K14/47C12N15/867
CPCC12N15/64C07K14/4748C07K14/705C12N15/86C12N2740/15043
Inventor 赵蔚
Owner 赵蔚
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