Recombinant single-chain antibody g5-4scfv of anti-human γδtcr monoclonal antibody and its encoding gene and application

Abstract

The invention discloses a recombinant single-chain antibody G5-4ScFv of an anti-human gamma delta T cell receptor (TCR) monoclonal antibody, and an encoding gene and the application of the recombinant single-chain antibody G5-4ScFv. A heavy chain variable region of the recombinant single-chain antibody of the anti-human gamma delta TCR monoclonal antibody has an amino acid residue sequence in a sequence table SEQ ID No.1, and a light chain variable region of the recombinant single-chain antibody of the anti-human gamma delta TCR monoclonal antibody has an amino acid residue sequence in a sequence table SEQ ID No.2. Experimental results show that the recombinant single-chain antibody G5-4ScFv can specifically bond with a gamma delta TCR; when the recombinant single-chain antibody G5-4ScFv and the gamma delta TCR are in a solid phase, the proliferation of gamma delta T cells in peripheral blood mononuclear cells (PBMC) of a human body can be stimulated, and the purity of the gamma delta T cells can be up to 90% in two weeks; and the gamma delta T cells obtained through the proliferation can effectively secrete interferon-gamma (IFN-gamma) and transforming growth factor-alpha (TGF-alpha) when being stimulated, and can effectively kill daudi cells which are in a tumor cell line, and therefore the recombinant single-chain antibody G5-4ScFv can be described to have a good cytotoxicity effect, and can be used in clinical adoptive immunotherapy to a tumor. Besides, the recombinant single-chain antibody G5-4ScFv has the following advantages in application: immunogenicity is lowered, and the safety of the clinical application is improved; the molecular weight is small, the penetration ability is excellent, and the recombinant single-chain antibody G5-4ScFv can easily enter the microenvironment around the solid tumor; and the recombinant single-chain antibody G5-4ScFv can be produced in a bulk, so that the output is increased, and the cost is lowered. The recombinant single-chain antibody G5-4ScFv plays an important role in the medical field, especially in the gamma delta T cell proliferation and tumor treatment, and has broad application prospects.

Description

technical field [0001] The present invention belongs to the monoclonal antibody and its encoding gene and application, in particular to the recombinant single-chain antibody G5-4ScFv (also referred to as "G5-4ScFv" herein) of the anti-human γδTCR monoclonal antibody and its encoding gene and application. Background technique [0002] T cells can be divided into two types according to the differential expression of surface receptors (TCR): TCRαβ cells and TCRγδ cells. γδT cells are a small subset of T lymphocytes, accounting for about 1%-5% of the total number of normal peripheral blood T lymphocytes, but they are relatively enriched in epithelial tissues such as epidermis and mucosa. In tumor immunity, the recognition of γδT cells to tumor antigens is not MHC-restricted and does not require specific antigen presentation, and has cytotoxic activity against a variety of tumor cells, so it has the potential to be used in tumor adoptive immunotherapy. Antibody solid-phase in vi...

AI Technical Summary

Problems solved by technology

Antibody solid-phase in vitro expansion is an important means to obtain γδT cells, but because the current anti-γδTCR monoclonal antibody is of mouse origin, it will produce human anti-mouse antibody reaction (HAMA), so it is greatly limited in clinical treatment

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