Antibody capable of combining interleukin 4 acceptor

A technology of interleukin and antibody, applied in the direction of fusion cells, antibodies, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc.

Active Publication Date: 2017-12-15
SUZHOU CONNECT BIOPHARMACEUTICALS LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunomodulatory therapy has achieved results in animal models, but human clinical trials still need to be explored

Method used

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  • Antibody capable of combining interleukin 4 acceptor
  • Antibody capable of combining interleukin 4 acceptor
  • Antibody capable of combining interleukin 4 acceptor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0176] Example 1 : Preparation of the antibody of the present invention and non-reducing SDS-PAGE gel electrophoresis to determine the expression level

[0177] The coding sequence of the antibody light chain variable region was inserted into the pFUSE2ss-CLIg-hK vector (Invivogen, product number: pfuse2ss-hclk) using EcoRI and BsiWI restriction sites to form a light chain expression vector; the coding sequence of the heavy chain variable region was used EcoRI and NheI restriction sites were inserted into pFUSEss-CHIg-hG2 vector (Invivogen, product number: pfusess-hchg2) or pFUSEss-CHIg-hG4 vector (Invivogen, product number: pfusess-hchg4) to constitute the heavy chain expression vector.

[0178] Expi293 cells were cultured and transfected according to Invitrogen Expi293 TM Expression SystemKit Manual (Cat. No. A14635). Cell density was adjusted to 2×10 for transfection 6 cells / ml, 0.6 μg light chain expression vector and 0.4 μg heavy chain expression vector were added to...

Embodiment 2

[0189] Example 2 : Detecting that the antibody of the present invention inhibits the proliferation of hIL-4 or hIL-13 on TF-1 cells

[0190] 1. Preparation of reagents

[0191] hIL-4 (Invivogen, product number: rhil-4) solution: dissolve hIL-4 with 100 μl of PBS solution containing 0.1% BSA (Beyotime, product number: ST023) to obtain a concentration of 100 μg / ml, and dissolve the dissolved hIL-4 according to A volume of 5 μl per tube was dispensed into 1.5ml (Nunc) centrifuge tubes and stored in a -20°C refrigerator.

[0192] WST-1 (Beyotime, product number: C0036) solution: Add 5ml of electronic couplant (C0036-2) to WST-1 powder (C0036-1), dissolve completely to form WST-1 solution, and divide according to the volume of 620μl per tube Pack into a 1.5ml centrifuge tube and store in a -20°C refrigerator.

[0193] 2. TF-1 cell culture

[0194] Take out the frozen TF-1 in liquid nitrogen (ATCC: CRL-2003 TM ) cells were shaken in a 37°C water bath to rapidly dissolve them. ...

Embodiment 3

[0215] Example 3 : ELISA detects the binding ability of the antibody of the present invention to sIL-4Rα

[0216] 1. Preparation of reagents

[0217] sIL-4Rα (PEPRO TECH, product number: 200-04R) solution: Take a tube of sIL-4Rα, add 1ml ddH2O, mix upside down to get a 100μg / ml solution, then store in -20℃ refrigerator after aliquoting.

[0218] Sample to be tested: get the Expi293 cell culture supernatant (the culture medium is Expi293Expression Medium, Invitrogen, article number: A1435102; in 8% CO2 incubator, continuous 100rpm suspension culture 4 days) 10 μ l and 2 μl was added to 990 μl and 998 μl of PBS solution respectively to prepare 1:100 and 1:500 diluted antibody samples to be tested.

[0219] Control sample: normal cell culture supernatant (Expi293 cells were not transfected, and the culture medium was Expi293Expression Medium (Invitrogen, article number: A1435102; in 8% CO2 incubator, continuous 100rpm suspension culture for 4 days) was also carried out at 1:10...

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Abstract

The invention provides an antibody capable of combining an interleukin 4(IL-4) acceptor (IL-4R). In addition, the invention provides a nucleic acid sequence for coding an antibody, a carrier containing the nucleic sequence and a host cell for converted or transfected by the carrier. Further, the invention provides a method for producing the antibody, a medical science application of the antibody and a kit containing the antibody.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular, the invention relates to an antibody capable of binding interleukin 4 (IL-4) receptor (IL-4R) and its application. Background technique [0002] Interleukin-4 (IL-4) is a cytokine mainly produced by activated T cells, monocytes, basophils, mast cells and eosinophils. IL-4 is involved in a variety of biological processes, and its known biological effects include stimulating the proliferation of activated B cells and T cells, CD4 + T cells differentiate into type II helper T cells. Studies have shown that IL-4 has multiple functions in mediating the immune response of allergic diseases, autoimmune diseases, infectious diseases, tumors and other diseases, and has therapeutic effects on tumors, autoimmune diseases and infectious diseases, and IL-4 has a regulatory effect on vaccine immune response, so IL-4 has always been one of the research hotspots that people pay attention to. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28A61K39/395A61P29/00A61P37/08
CPCC07K16/2866C07K2317/92C07K2317/76C07K2317/567C07K2317/565C07K2317/24A61K47/6849A61K45/06A61P29/00A61P37/08A61P17/00A61P11/06A61P1/00A61P11/02A61P19/02C07K2317/56A61K2039/505A61K39/395C07K16/28A61K38/00C12N15/62A61P39/00C07K16/247C12N5/12C12N15/85C12N2015/8518
Inventor 郑伟潘武宾杨欣陈阳张立民蒋洁
Owner SUZHOU CONNECT BIOPHARMACEUTICALS LTD
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