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Cytoplasma membrane surface antigen monoclonal antibody and uses thereof

A monoclonal antibody and surface antigen technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, anti-tumor drugs, etc., can solve the problems of undisclosed esophageal cancer metastasis monoclonal antibodies, and achieve the effect of inhibiting metastasis

Inactive Publication Date: 2011-10-12
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most importantly, there are no biological products and monoclonal antibodies for the prevention and treatment of esophageal cancer metastasis at home and abroad.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of Mouse Monoclonal Antibody Binding to Human Esophageal Cancer Cell Membrane Surface Antigen

[0031] Esophageal cancer cells were used to immunize Bal b / c mice to prepare monoclonal antibodies against esophageal cancer cells, and live cell immunofluorescence was used to screen the mouse monoclonal antibodies that combined with antigens on the membrane surface of human esophageal cancer cells. Specifically, a single esophageal cancer cell that was ground and separated from esophageal cancer tissue fixed with paraformaldehyde was used to immunize Bal b / c mice, subcutaneously immunized once every 3 weeks, and the tail blood of the mice was taken to detect the degree of antiserum binding to esophageal cancer cells. Titer titer until the titer reaches 1; after 50,000, mouse splenocytes are fused with SP2 / 0 cells according to the conventional PEG fusion method to prepare monoclonal antibody. Hybridoma supernatants were collected using 96-well culture p...

Embodiment 2

[0032] Example 2 Screening and identification of monoclonal antibodies that inhibit the growth of human esophageal cancer cells

[0033] MTT, MTS, CCK-8, etc. were used to determine the effect of monoclonal antibody culture supernatant on the proliferation of esophageal cancer cells. Specifically, the MTT method was used to inoculate esophageal cancer cell Yes2 into a 96-well culture plate at a density of 2500 cells / well. After the cells adhered to the wall for 12 hours, the culture medium was replaced with 1:2 diluted ECS6C5 monoclonal antibody culture supernatant. After the cells were cultured for 72 hours, discard the supernatant, add 5mg / ml MTT solution, incubate at 37°C for 4 hours, discard the solution, add 150ul DMSO to dissolve formazan, and measure the absorbance at 570nm. The measured value of the supernatant of the ECS6C5 monoclonal antibody was 81.50%±11.12% of the measured value of the SP2 / 0 supernatant control group, and there was a statistically significant diff...

Embodiment 3

[0034] Example 3 Screening and identification of monoclonal antibodies that inhibit the adhesion of human esophageal cancer cells to microvascular endothelial cells derived from human esophageal cancer tissue

[0035] The effect of monoclonal antibody culture supernatant on the adhesion of esophageal cancer cells to microvascular endothelial cells derived from human esophageal cancer tissue was determined by metabolic fluorescent labeling method. Specifically, esophageal cancer tissue-derived microvascular endothelial cells ECEC were inoculated into gelatin-coated 96-well culture plates at a density of 9,000 cells / well. Endothelial resting medium with 0.1% FBS. After the tumor cells were labeled with Calcein AM, they were incubated with the culture supernatant of ECS6C5 monoclonal antibody at 37 degrees for 1 hour, and then added to the static endothelial monolayer at 50,000 cells / well at 37 degrees in 5% CO 2 After 1 hour of incubation, the 1640 medium was washed 4 times. A...

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Abstract

The present invention relates to a novel monoclonal antibody which can be combined with the natural membrane antigen of the human esophageal cancer cells, the hybridoma series of the monoclonal antibody as well as the application of the monoclonal antibody in the preparation of drugs that can control the transfer of human esophageal cancer cells. The monoclonal antibody can be combined with the natural antigens on the membrane surface of the EC-9706 cells, human esophageal cancer cells that are of transfer capability, as well as the natural antigens on the membrane surface of the human esophageal cancer cells YES2. The monoclonal antibody can effectively control the proliferation, migration, adhesion and invasion of human esophageal cancer cells in vitro and can effectively restrain the transfer of human esophageal cancer cells in vivo. The present invention also provides the production series mouse hybridoma cells series of the monoclonal antibody. The preservation number is CGMCC No.1723.

Description

technical field [0001] The present invention relates to an antibody capable of binding to human cancer cells and the hybridoma cell line produced therefrom, as well as the application of the antibody in treating cancer metastasis. Specifically, the present invention provides a monoclonal antibody that can bind to the natural membrane antigen of human esophageal cancer cells, and its application in the preparation of drugs that can be used to treat human esophageal cancer metastasis. Background technique [0002] Esophageal cancer is one of the common human tumors with high morbidity and mortality. At present, the five-year mortality rate of esophageal cancer is still above 90%, which seriously threatens human life and health. The important reason for the high mortality rate of esophageal cancer is that esophageal cancer is different from other malignant tumors. Metastasis is not only early but also fast. In many cases, metastasis has already occurred when the primary tumor r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N5/18A61K39/395A61P35/00
Inventor 杨治华冉宇靓胡海
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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