Bispecific antibody FITC*CD3 as well as preparation method and application thereof

A bispecific antibody and specific technology, applied in the field of bispecific antibody FITC×CD3 and its preparation, can solve the problems of cumbersome purification steps, difficult prokaryotic expression, poor stability, etc., and achieve uniform properties, high binding capacity and stability. good effect

Active Publication Date: 2018-04-17
康梦实
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problems of existing bispecific antibodies such as difficulty in prokaryotic expression, low yield, cumbersome purification steps, and poor stability, the present invention provides a bispecific antibody FITC×CD3 and its preparation method and application

Method used

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  • Bispecific antibody FITC*CD3 as well as preparation method and application thereof
  • Bispecific antibody FITC*CD3 as well as preparation method and application thereof
  • Bispecific antibody FITC*CD3 as well as preparation method and application thereof

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Embodiment 1

[0034] 1. Design of bispecific antibodies

[0035] The bispecific antibody targeting FITC and CD3 is named FITC×CD3, and the structure of the antibody can be found in figure 1 , one of the monovalent antigens is an anti-FITC heavy chain-light chain pair (SEQ ID NO: 1), and the other monovalent antigen is an anti-CD3 heavy chain-light chain pair (SEQ ID NO: 2), and the sequence refers to a monoclonal antibody The sequence of L2K (refer to US20070123479 sequence number 2), the two monovalent antigens are connected by the Fc segment of human immunoglobulin 1 (SEQ ID NO: 3), and the Fc segment is connected to the anti-CD3 heavy chain-light chain A flexible linker sequence (SEQ ID NO: 4) was added between them. In order to enable FITC×CD3 to be expressed in Chinese hamster ovary cells (CHO) cells and secreted into the medium, the leader peptide sequence of the mouse immunoglobulin κ light chain was selected as the secretion signal peptide. The signal peptide is directly linked to...

Embodiment 2

[0061] Pharmacodynamic study of bispecific antibody against tumor

[0062] Mix equal volumes of phosphate buffered saline (PBS) and peripheral blood of healthy people, add equal volumes of lymphocyte separation medium, centrifuge at room temperature for 30 minutes at 2500 rpm. Take the middle buffy coat, add 10 times the volume of PBS to wash away the lymphocyte separation solution, repeat twice, count and resuspend in RPMI 1640 complete medium to obtain peripheral blood mononuclear cells (PBMC). Cell viability >95% was confirmed by trypan blue staining and cell counts were performed. Separated PBMC cells were added to the cultured tumor cell culture medium at the same time, so that the ratio of PBMC cells to tumor cells was 5:1, and the mixed cultured cells were divided into 6 groups. Group 1 was regarded as a negative group without drug addition, and group 1 2 Add 1 μg of FITC-modified antibody bevacizumab, add 1 μg of the bispecific antibody of the present invention and 1 ...

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Abstract

The invention discloses a bispecific antibody FITC*CD3 as well as a preparation method and an application thereof and belongs to the fields of biological pharmacy and immunology. An immune cell antigen targeted by the bispecific antibody FITC*CD3 is CD3, and a targeted target antigen is fluorescein FITC. After the bispecific antibody FITC*CD3 is combined with an FITC modified antibody, T-lymphocytes can be targeted to tumor cells to kill the tumor cells. Compared with independent use of the antibody, the bispecific antibody can enhance pharmaceutical effect and promote immunoreaction of an organism for the tumor cells. The bispecific antibody FITC*CD3 is combined with different FITC modified antibodies, so that the obtained bispecific antibody can target multiple kinds of tumor cells. Theantibody is a biomacromolecular treatment drug prepared with a cell engineering technology and a gene engineering technology and has the advantages of being high in specificity, uniform in property, capable of realizing directional preparation by aiming at specific target spots and the like.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals and immunology, and more specifically relates to a bispecific antibody FITC×CD3 and its preparation method and application. Background technique [0002] Bispecific antibodies are genetically engineered antibodies containing two antigen-binding sites, which can build bridges between target cells and functional cells to produce specific effector functions. In biomedicine, especially in tumor immunotherapy, it has a very broad application. Killing tumor cells through bispecific antibody-mediated cytotoxicity is a hot spot in current immunotherapy application research, and its main feature is that bispecific antibodies can simultaneously bind to specific antigens on the surface of tumor cells and target antigens on immune effector cells, directly Stimulate the specific killing effect of immune effector cells on tumor cells. However, in the process of bispecific antibody drug development,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/46C12N15/13C12N15/85A61K39/395A61P35/00A61P35/02
CPCA61K2039/505C07K16/468C07K2317/56
Inventor 郭长缨康梦实
Owner 康梦实
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