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A fully human her2 antibody, its coding gene and application

A fully human, antibody technology, applied in applications, antibodies, genetic engineering, etc., can solve problems such as unmet demand and recurrence

Active Publication Date: 2018-02-23
GENOR BIOPHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although multiple options are available for the treatment of HER2-positive metastatic breast cancer, there is still an unmet need
Optimal regimens for the disease continue to emerge, but most patients with metastatic breast cancer relapse

Method used

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  • A fully human her2 antibody, its coding gene and application
  • A fully human her2 antibody, its coding gene and application
  • A fully human her2 antibody, its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening clones that specifically bind to human HER2-Fc from a fully human scFv phage library

[0043] Using the antigen-antibody binding specificity of ELISA technology, the human HER2 (extracellular domain)-Fc fusion protein (referred to as hHER2-Fc) antigen is coated on the microtiter plate, and it is specifically adhered to the coating by washing and panning. Phage on antigen. PBS (0.01M Na 2 HPO 4 12H 2 O+0.002M KH 2 PO 4 +0.14M NaCl+0.002M KCl, pH=8.6) was diluted to 5 μg / ml, added to a microtiter plate at 100 μl / well, and coated overnight at 4°C. After the plate was washed 4 times with PBST (PBS buffer containing 0.05% Tween 20), 300 μl / well of 5% BSA (purchased from Amresco, product number: 0332-100g, solution in PBS) was added, and blocked for 1 hour at 37°C. Then wash the plate twice with PBST. will contain 7×10 10 Independently cloned fully human scFv phage antibody library (this antibody library was constructed by Eureka (Beijing) Biotechn...

Embodiment 2

[0044] Example 2 Using enzyme-linked immunosorbent assay (ELISA) to identify the immunoreactivity of phages specifically binding to human HER2-Fc

[0045]The immunoreactivity of the phage specifically binding to human HER2-Fc obtained in Example 1 was further identified by enzyme-linked immunosorbent assay (ELISA). Human HER2-Fc antigen (purchased from Sino Biological Company, product number: 10004-H02H) was diluted 2 μg / ml with PBS (pH=8.6), added to the microtiter plate at 100 μl / well, and coated overnight at 4°C. After washing the plate 4 times with PBST, 300 μl / well of 5% BSA (purchased from Amresco, product number: 0332-100g, PBS solution) was added, and blocked at 37°C for 1 hour. Then wash the plate twice with PBST, add 100 μl / well phage clone suspension, and incubate at 37° C. for 2 hours. Wash the plate 4 times with PBST. Add HRP-labeled anti-M13K07 phage antibody (GE, catalog number: 27-9421-01, diluted 1:5000 in PBST, 100 μl / well), and incubate at room temperature...

Embodiment 3

[0049] Example 3 Detecting Species Cross-reactivity of 102 Human HER2-Fc-specific scFvs and Cross-reactions Between HER Family Member Molecules by ELISA

[0050] The species cross-reactivity of 102 human HER2-Fc-specific scFv and cross-reaction with HER family members were detected by ELISA. The method was the same as in Example 2, except that the coated human HER2-Fc antigen was replaced with monkey HER2-Fc (purchased from Sino Biological Company, product number: 90295-C02H), mouse HER2-Fc (purchased from Sino Biological Company, product number: 50714 -M02H), human HER1-Fc (purchased from Sino Biological Company, product number: 10001-H02H), human HER3-Fc (purchased from Sino Biological Company, product number: 10201-H05H) and human HER4-Fc (purchased from Sino Biological Company , Item No.: 10363-H02H). The antigen was diluted 2 μg / ml with PBS (pH=8.6), added to the microtiter plate at 100 μl / well, and coated overnight at 4°C. After washing the plate 4 times with PBST, 300...

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Abstract

The invention relates to the field of medicinal chemistry, in particular to two fully human HER2 antibodies, their coding genes and applications. The present invention provides two fully human HER2 antibodies, wherein the amino acid sequence of the heavy chain variable region of one HER2 antibody is SEQ ID NO: 1, the amino acid sequence of the light chain variable region is SEQ ID NO: 2, and the other The amino acid sequence of the heavy chain variable region of the HER2 antibody is SEQ ID NO:10, and the amino acid sequence of the light chain variable region is SEQ ID NO:11. The fully human HER2 antibody can reduce infusion reactions and immunogenicity, improve drug safety, and have better pharmacokinetic characteristics. In addition, the fully human antibody of the present invention can be used in combination with other HER2-positive tumor therapeutic agents to treat HER2-positive tumors.

Description

technical field [0001] The invention relates to the technical field of antibodies, in particular to a fully human HER2 antibody, its coding gene and its application. Background technique [0002] Breast cancer is the most common malignancy in women worldwide. The HER family regulates the growth and development of normal mammary glands, and overexpression of HER2 is associated with breast cancer. Trastuzumab (trastuzumab), trade name Herceptin (Herceptin), is the first humanized monoclonal antibody for the treatment of human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer drug. Although Herceptin has become the standard treatment for HER2-positive malignancies, 40% of patients still do not respond to Herceptin. Furthermore, drug resistance has become a common and serious problem in anti-HER2 therapeutics. Redundancy of growth factors and crosstalk between intracellular signaling pathways are thought to be major contributors to drug resistance i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/30C12N15/13A61K39/395A61P35/00A61P35/02G01N33/68G01N33/574
Inventor 周清舒孟军石姝言苏谭涛超
Owner GENOR BIOPHARMA
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