Anti-cd3 antibody, and molecule containing said antibody

A technology of antibody and CDRH2, applied in the direction of antibody, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, etc., can solve problems such as treatment drug barriers

Pending Publication Date: 2019-11-08
DAIICHI SANKYO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 7) A limitation of OKT3 and a series of modified OKT3 antibodies is their specificity for human CD3
This limitation may pose a considerable obstacle to the development of therapeutic drugs for the treatment of human diseases

Method used

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  • Anti-cd3 antibody, and molecule containing said antibody
  • Anti-cd3 antibody, and molecule containing said antibody
  • Anti-cd3 antibody, and molecule containing said antibody

Examples

Experimental program
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Embodiment 1）-7

[0397] Examples of chimeric antibodies include, but are not limited to, antibodies comprising a non-human animal antibody-derived variable region joined to a human antibody (human immunoglobulin) constant region.

[0398] Examples of humanized antibodies may include, but are not limited to, human antibodies grafted with CDRs in variable regions of non-human animal antibodies (human immunoglobulin variable regions), CDRs grafted in non-human animal antibodies, and framework regions. Partial sequences of human antibodies, and antibodies having human antibody amino acids substituted for one or more amino acids derived from non-human animal antibodies in any of these humanized antibodies. Examples of the CDRs in the variable region of the non-human animal antibody include CDRH1 to CDRH3 in the heavy chain variable region and CDRL1 to CDRL3 in the light chain variable region derived from the rat anti-CD3 antibody C3E-7000 of the present invention, And CDRs derived from the amino ac...

Embodiment 1）

[0667] (Example 1) Preparation of rat anti-human CD3 antibody

[0668] 1) Construction of -1 human CD3εδ expression vector

[0669] The control vector pcDNA3.1-DEST transformed into the destination vector was prepared using Gateway Vector Conversion System (Thermo Fisher Scientific Inc.). figure 1 The cDNA (SEQ ID NO: 1) encoding the human CD3ε protein (NCBI reference sequence: NP_000724.1) shown in was purchased from Sino Biological Inc., and cloned into pcDNA3 using Gateway LRClonase Enzyme mix (Thermo Fisher Scientific Inc.). 1-DEST vector to construct hCD3ε-pcDNA3.1. Using human T cell-derived cDNA as a template, the encoding figure 2 The cDNA of the human CD3δ protein (NP_000723.1) (SEQ ID NO: 2) shown in and cloned in pcDNA3.1(+) (Thermo Fisher Scientific Inc.) to construct the expression vector hCD3δ-pcDNA3.1. For large-scale preparation of each expression vector, Endofree Plasmid Giga Kit (Qiagen N.V.) was used.

[0670] 1) -2 immunity

[0671] For immunization, ...

Embodiment 2）

[0701] (Example 2) Study on Binding Activity of Rat Anti-CD3 Monoclonal Antibody (C3-147) to Human CD3

[0702] 2)-1 Preparation of monoclonal antibody from hybridoma supernatant

[0703] 2)-1-1 Cultivation of C3-147-producing hybridomas

[0704] Rat anti-CD3 monoclonal antibody was purified from hybridoma culture supernatant. First, C3-147-producing hybridomas were allowed to grow to sufficient quantities in ClonaCell-HY Selection Medium E (StemCell Technologies Inc.). Then, the medium was replaced with Hybridoma SFM (Thermo Fisher Scientific Inc.) supplemented with 20% UltraLow ​​IgG FBS (Thermo Fisher Scientific Inc.) containing 5 μg / mL gentamicin (Thermo Fisher Scientific Inc. .), followed by culturing for 7 days. The culture supernatant was recovered and sterilized by passing through a 0.22 μm filter (Corning Inc.).

[0705] 2) -1-2 purification

[0706] Antibodies were purified from the hybridoma culture supernatant prepared in Example 2)-1-1 by protein G affinity c...

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Abstract

The present invention provides a novel antibody capable of binding to human CD3; or a molecule which is capable of binding to an antigen and contains the antibody. Provided are a novel antibody capable of binding to human CD3; a molecule which contains the antibody and is capable of binding to an antigen; a pharmaceutical composition which contains the antibody or the molecule as an active ingredient and has a cytotoxic activity; and others.

Description

[technical field] [0001] The present invention relates to novel antibodies that bind to human CD3 and to cynomolgus monkey CD3, and molecules comprising said antibodies. [Background technique] [0002] 1) Like any other monoclonal antibody, an anti-CD3 monoclonal antibody works by precisely recognizing its target molecule. Each anti-CD3 antibody recognizes only a single epitope on the target CD3 molecule. Among the monoclonal antibodies specific for the CD3 complex, OKT3 is the most widely used and best characterized. [0003] 2) OKT3 is a mouse-derived anti-human CD3 monoclonal antibody (Non-Patent Document 1). Anti-CD3 monoclonal antibodies have been administered intraoperatively to prevent rejection of transplanted organs. The antibody binds to the TCR complex on human T cells and inhibits their activation and proliferation. This treatment has been used for a long time in order to prevent allograft rejection of organs (Non-Patent Documents 2 to 4). OKT3 is the first ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K39/395A61P43/00C07K16/28C12N1/15C12N1/19C12N1/21C12N5/10
CPCC12N15/09A61K2039/505C07K2317/24C07K2317/31C07K2317/33C07K2317/56C07K2317/622C07K2317/92C07K16/2809C07K16/2827C07K16/2863C07K16/30C07K2317/34C07K2317/73A61P35/00C07K2317/52C07K2317/732C07K16/32C07K2317/53C12N15/85C12N2015/8518
Inventor 高桥亘吉村千草上妻志保中村健介铃木智香子市川淳也
Owner DAIICHI SANKYO CO LTD
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