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Monoclonal antibody and application thereof

A monoclonal antibody, sequence technology, applied in application, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, etc., can solve the lack of costimulatory signal, T cells Clonal anergy, inability to activate T cells, etc.

Active Publication Date: 2010-01-20
广东云天抗体生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The lack of co-stimulatory signals during the induction phase of T cell activation not only fails to activate T cells, but also causes T cell clone-specific anergy, leading to immune tolerance

Method used

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  • Monoclonal antibody and application thereof
  • Monoclonal antibody and application thereof

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Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1, the preparation of monoclonal antibody anti-4-1BB-CTLA4-IgG2a

[0024] 1. Obtaining the heavy chain and light chain of monoclonal antibody anti-4-1BB-CTLA4-IgG2a

[0025] The genes encoding the heavy chain variable regions of the anti-CTLA-4 antibody and the anti-4-1BB antibody were artificially synthesized, respectively. The obtained anti-CTLA-4 antibody and the coding gene of the heavy chain variable region of the anti-4-1BB antibody are connected by overlapping extension PCR to obtain the DNA sequence of the heavy chain variable region of the monoclonal antibody. The amino acid residue sequence encoded by the DNA sequence of the chain variable region is shown in Sequence 1.

[0026] The coding gene of the variable region of the heavy chain of the monoclonal antibody obtained above and the coding gene of the constant region of the isotype mouse IgG2a are respectively connected by overlapping extension PCR to obtain the DNA sequence of the heavy chain of...

Embodiment 2

[0034] Example 2, Identification of binding ability of monoclonal antibody anti-4-1BB-CTLA4-IgG2a

[0035] 1. Identification of the binding ability of monoclonal antibody anti-4-1BB-CTLA4-IgG2a to CTLA-4

[0036] The plate was coated with CTLA-4-Fc monoclonal antibody (purchased from R&D Company, product number: 325-CT), and each well was coated with 1 ug / ml. The monoclonal antibody anti-4-1BB-CTLA4-IgG2a constructed in Example 1 was serially diluted and incubated, and the secondary antibody was goat anti-mouse IgG labeled with horseradish peroxidase (purchased from Jackson ImmunoResearch, catalog number: 115-035-062) , with OPD as substrate and 2mmol / L sulfuric acid as terminator, ELISA was used to detect the binding ability of monoclonal antibody anti-4-1BB-CTLA4-IgG2a to CTLA-4. The result is as figure 1 as shown, figure 1 In , the ordinate is the light absorption value after ELISA color development. The results showed that the monoclonal antibody anti-4-1BB-CTLA4-IgG2a...

Embodiment 3

[0039] Embodiment 3, the preparation of the Fab fragment of monoclonal antibody anti-4-1BB-CTLA4-IgG2a

[0040] use Monoclonal antibody anti-4-1BB-CTLA4-IgG2a secreted by transiently transfected mammalian COS-7 cells was digested with immobilized papain in the Fab Preparation Kit (Pierce) to degrade the full-length mAb into Fab and Fc fragment. The digested product was purified with the immobilized protein A column provided in the kit to obtain Fab fragments. ELISA identification was carried out according to the method of Example 2. The results showed that the Fab fragments had comparable binding ability to full-length antibodies.

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Abstract

The invention discloses a monoclonal antibody and application thereof. The monoclonal antibody consists of light chains and heavy chains, wherein amino acid residue sequences of the variable area of the heavy chains are showed in a sequence 1 in the sequence table, and amino acid residue sequences of the variable area of the light chains are showed in a sequence 2 in the sequence table. The invention combines and constructs anti-CTLA-4 antibodies and anti-4-1BB antibodies to form antihuman 4-1BB and CTLA-4 monoclonal antibodies. Experimental result shows that the monoclonal antibody has an antibody structure and can be respectively combined with human 4-1BB and CTLA-4.

Description

technical field [0001] The present invention relates to a monoclonal antibody and its application. Background technique [0002] Theoretically speaking, a large number of cells in the human body mutate every day, but only a very small number of mutated cells can continue to divide and grow, and escape the surveillance and attack of the immune system to form tumors. Tumors originate from the body's own mutated cells. Most of the mutated cells have the same components as the body's normal cells, and only a small number of abnormally expressed proteins and deformed polysaccharides are immunogenic. Due to the difference in immunogenicity among tumor cells, those tumors with strong immunogenicity can induce effective anti-tumor immune response and are easily eliminated by the body; while those tumors with relatively weak immunogenicity can escape the immune system This process is called immune selection; after continuous selection, the immunogenicity of tumors becomes weaker and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N15/13C12N15/63C12N5/10C12N1/00
Inventor 唐捷刘阳陈列平朱卫彬
Owner 广东云天抗体生物科技有限公司
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