Antibody binding to LAG3 and application of antibody

An antibody and monoclonal antibody technology, used in infectious diseases and autoimmune diseases, to treat LAG3-related diseases such as cancer, and to solve problems such as the occurrence and deterioration of autoimmune diseases

Active Publication Date: 2020-10-23
BEIJING MABWORKS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, LAG3 deficiency may trigger the development and exacerbation of autoimmune diseases

Method used

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  • Antibody binding to LAG3 and application of antibody
  • Antibody binding to LAG3 and application of antibody
  • Antibody binding to LAG3 and application of antibody

Examples

Experimental program
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preparation example Construction

[0125] Preparation of the monoclonal antibody of this application

[0126] The monoclonal antibodies of the present application can be prepared using the somatic cell hybridization (hybridoma) technique of Kohler and Milstein (1975) Nature 256:495. Other embodiments for producing monoclonal antibodies include viral or oncogenic transformation of B lymphocytes and phage display techniques. Chimeric or humanized antibodies are also well known in the art. See, eg, US Patents 4,816,567; 5,225,539; 5,530,101; 5,585,089; 5,693,762 and 6,180,370.

[0127] Generation of transfectomas for the preparation of the monoclonal antibodies of the present application

[0128] Antibodies of the present application can also be produced in host cell transfectomas using, for example, recombinant DNA technology combined with gene transfection methods (eg Morrison, S. (1985) Science 229:1202). In one embodiment, DNA encoding partial or full-length light and heavy chains obtained by standard m...

Embodiment 1

[0169] Example 1 Construction of HEK293A cell strain stably expressing human, monkey or mouse LAG3

[0170] HEK293A cells were used to construct cell lines stably overexpressing human, monkey or mouse LAG3. Briefly, the cDNA sequences of human, monkey or mouse LAG3 (SEQ ID NOs: 37, 38 and 39) were synthesized and cloned into the pLV-EGFP(2A)-Puro vector (Beijing Yingmaoshengye Biotechnology Co., Ltd. company, China) between EcoRI and XhoI restriction sites. The resulting pLV-EGFP(2A)-Puro-VISTA, psPAX and pMD2.G plasmids were transfected into HEK293T cells (Nanjing Kebai Company, China) by liposome transfection to produce lentiviruses. The method was completely consistent with the steps in the instructions of the Lipofectamine 3000 kit (Thermo Fisher Scientific, USA). Three days after transfection, lentivirus was harvested from the cell culture medium (DMEM medium (Cat#: SH30022.01, Gibco) supplemented with 10% FBS (Cat#: FND500, Excell)) of HEK293T cells. Then transfect ...

Embodiment 2

[0171] Example 2 Preparation of hybridoma cell lines producing mouse anti-human LAG3 monoclonal antibody

[0172] The mouse anti-human LAG3 monoclonal antibody was obtained by conventional hybridoma fusion technology, and the protocol was slightly modified, as follows.

[0173] immunity

[0174] Table 2. Immunization scheme

[0175]

[0176] Thirteen BALB / c mice (Victoria, China) were cross-injected with recombinant human LAG3(ECD)-hFc protein (Cat#: 16498-H02H, Sino Biological, China) and monkey LAG3(ECD)-hFc protein (Cat#: LA3-C5252, ACRO, China) for immunization, the specific immunization process is shown in Table 2. Human LAG3(ECD)-hFc protein and monkey LAG3(ECD)-hFc protein were treated with equal volume of complete Freund's adjuvant (Cat#: F5881-10*10ML, Sigma, USA) or incomplete Freund's adjuvant (Cat#: F5506 -6*10ML, Sigma, USA) or PBS phacoemulsification.

[0177] One week after each booster immunization, 50 μl of serum was collected from the tail of each mo...

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Abstract

The present application provides an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human LAG3. Also provided are a nucleic acid molecule encoding the antibody, as well as an expression vectors, a host cell, and a method for expressing the antibody. The application also provides an immune crosslinker, a bispecific molecule, a chimeric antigen receptor, and a pharmaceutical composition comprising the antibody, as well as a treatment method using the antibody provided by the application.

Description

technical field [0001] The present application relates to an antibody specifically binding to LAG3, its preparation and use, especially its use in the treatment of LAG3-related diseases such as cancer, infectious diseases and autoimmune diseases. Background technique [0002] The process of immune response is very complicated, and various stimulatory and / or inhibitory molecules on immune cells are activated or deactivated to maintain the immune response in an appropriate or optimal state. For example, inhibitory receptors such as PD-1 and CTLA-4 are upregulated to balance the activity of stimulatory receptors and limit the activation of immune cells, thus preventing the occurrence of autoimmunity or autoinflammation. However, this immunosuppressive mechanism is exploited by tumor cells to protect themselves from the immune system's attack, causing cancer initiation and progression. In the infection of foreign pathogens, especially in the case of chronic infection, LAG3 is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C07K14/725C07K19/00C12N15/13A61K45/06A61K39/395A61K38/20A61P35/00A61P31/00A61P37/02A61P31/12A61P31/04A61P31/10A61P17/06
CPCC07K16/2803C07K14/7051A61K45/06A61P35/00A61P31/00A61P37/02A61P31/12A61P31/04A61P31/10A61P17/06C07K2317/33C07K2317/92C07K2317/24C07K2317/56C07K2317/565A61K2039/505A61K2039/507C07K2317/76C07K2317/73C07K16/2818A61K38/00A61K38/20A61K38/2013A61K39/39541A61K39/3955A61K39/39558C07K2317/21C07K2317/74
Inventor 胡稳奇李江美李锋
Owner BEIJING MABWORKS BIOTECH
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