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Monoclonal antibody for human ADRB3 (beta3 adrenoceptor) and application of monoclonal antibody to disease diagnosis and treatment

A monoclonal antibody, disease technology, applied in the field of biomedicine

Active Publication Date: 2017-10-20
郑猛 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] And for ADRB3 in malignant tumors, atherosclerosis, Alzheimer's disease, inflammation, sepsis, multiple organ dysfunction syndrome, viral infection, autoimmune disease, organ transplant immune rejection, aging, osteoporosis , severe trauma, poisoning, cachexia and other diseases, there are no relevant studies and reports

Method used

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  • Monoclonal antibody for human ADRB3 (beta3 adrenoceptor) and application of monoclonal antibody to disease diagnosis and treatment
  • Monoclonal antibody for human ADRB3 (beta3 adrenoceptor) and application of monoclonal antibody to disease diagnosis and treatment
  • Monoclonal antibody for human ADRB3 (beta3 adrenoceptor) and application of monoclonal antibody to disease diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0305] Example 1 Immunofluorescence detection of ADRB3 localization in tumor cells

[0306] 1. Experimental method

[0307] Breast cancer MCF7 cells, lung cancer A549 cells, melanoma A375 cells, pancreatic cancer CFPAC1 cells, liver cancer HepG2 cells, glioma A172 cells in 10 5Each well was inoculated in a 6-well plate with a built-in sterile cover glass, incubated until the cells were patched, fixed with 4% paraformaldehyde for 10 min, and washed 3 times with PBS. Permeabilize with 0.1% TritonX-100 for 10 minutes, block with 3% BSA for 1 hour, add antibodies such as Ki-67 / ADRB3 / Nucleolin / Fibrillarin / H3K9AC / Brdu / α-tubulin / Flag (1:100) dropwise, and use Mitotracker as the mitochondrial dye , Lysotracker was used for lysosome dye, Brdu was given before cell fixation, and lipofectin3000 was used to transfect pcDNA3-flag-B3 plasmid into cells. The wet box was sealed and incubated overnight in a 4°C refrigerator. Rinse with PBS, 5min×3, add FITC-labeled secondary antibody (anti-...

Embodiment 2

[0321] Example 2 Tissue chip immunohistochemical method to detect the expression of ADRB3 in tumor tissue

[0322] 1. In this study, tumor tissue chips from 11 different tissue sources were used, with a total of 1479 patients: (1) 42 cases of non-small cell lung cancer tissues; (2) 150 cases of lung adenocarcinoma with survival data; (3) 150 cases Survival lung squamous cell carcinoma; (4) 180 survival cases of liver cancer; (5) 170 survival cases of breast cancer and 82 cases of non-survival breast cancer; (6) 170 survival cases of pancreatic cancer; (7) 62 cases of thyroid papilla (8) 90 survival cases of colon cancer; (9) 90 survival cases of kidney cancer; (10) 93 survival cases of esophageal cancer; (11) 90 survival cases of gastric cancer; (12) 90 survival cases of rectal cancer Cancer; (13) 20 cases of leukemia.

[0323] 2. Experimental steps: (1) Paraffin section: thickness 4μm; (2) Baked slice: 65°C, 3h; (3) Dewaxing to water: xylene 10min×3 cylinders, 100%-100%-95%-...

Embodiment 3

[0346] Example 3 Immunofluorescence detection of ADRB3 expression in bone marrow smear cells of patients with different types of leukemia

[0347] 1. The experimental method is the same as in Example 1.

[0348] 2. The results are attached Figures 27 to 33 Shown:

[0349] (1) Myeloperoxidase (MPO) is a marker protein of neutrophils, and ADRB3 is expressed in normal human neutrophils, but not in lymphocytes. ADRB3 in the cytoplasm of neutrophils forms a conical structure to reach the nucleus, and the channel formed by ADRB3 helps extracellular substances (such as viruses) enter the nucleus, blocking ADRB3 will inhibit the virus from entering the nucleus. In B lymphocytic leukemia patients, ADRB3 is expressed in both leukemic cells and neutrophils, suggesting that ADRB3 induces carcinogenesis of B lymphocytes. ADRB3 in leukemia cells also forms channels to the nucleus

[0350] (2) ADRB3 and MPO staining of normal human bone marrow smear after injection of Granoside (recombi...

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Abstract

The invention discloses an application of ADRB3 (beta3 adrenoceptor) as a marker for detecting various diseases and an application of a monoclonal antibody for human ADRB3 to preparation of disease diagnosis and treatment drugs. ADRB3 is involved in and participates in various signaling pathways and can serve as a diagnostic marker and a diagnostic target for various diseases; the monoclonal antibody for the human ADRB3 has the functions of resisting cancer, inflammation, viruses, shock, allergy, virus infectious diseases, autoimmune diseases, diseases caused by regenerative dysfunction, cachexia, cardiovascular and cerebrovascular diseases, neurodegenerative diseases and aging, the function of autophagy regulation and other functions and has important medical value and research and application prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the technology of immune regulation signal pathway. More specifically, the anti-human β3 adrenergic receptor monoclonal antibody is involved in the preparation of anti-cancer, anti-inflammation, anti-drug, anti-shock, anti-allergy, anti-viral infection, anti-autoimmune disease, anti-cachexia, anti-atherosclerosis , new uses in anti-neurodegeneration, regulation of autophagy and anti-aging drugs. Background technique [0002] The β3 adrenoceptor (Adrenoceptor Beta 3, ADRB3, B3AR, beta3adrenoceptor, β3 receptor) in the human body is a type of tissue receptor that mediates the action of catecholamines, and it is a G protein-coupled type. The current research on ADRB3 mainly focuses on the effects on body fat, blood sugar and blood lipid levels, insulin secretion and action. [0003] And for ADRB3 in malignant tumors, atherosclerosis, Alzheimer's disease, inflammati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/68C07K14/72C07K16/28C12N5/20C12R1/91
CPCC07K14/70571C07K16/286C07K16/2869G01N33/577G01N33/68C07K2317/565C07K14/705A61K2039/505G01N33/74G01N33/9433G01N2333/726A61P25/36A61P35/00A61P37/00A61P25/18A61P15/10A61P31/12A61P3/10C07K2317/24C07K2317/622C07K2317/73C07K2319/03C07K2319/33G01N33/56966G01N33/56972C07K14/7051G01N33/57484G01N2333/70571A61K39/395C07K14/70517C07K14/70578C07K2319/00G01N33/6893G01N33/6896
Inventor 郑猛林曙光
Owner 郑猛
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