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Monoclonal antibody resisting ovarian cancer and application thereof

A monoclonal antibody and ovarian cancer technology, applied in the field of medical diagnosis, can solve the problems of patients losing radical surgery, atypical signs, and low specificity

Active Publication Date: 2016-04-06
潘世扬
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the anatomical position of the ovary deep in the pelvic cavity, the onset is hidden, and the early clinical symptoms and signs are not typical. About 70% of the patients are already in the advanced stage when they see a doctor, and most of them have metastasized, resulting in most patients losing the chance of radical surgery, leading to five The annual survival rate is low, less than 30%
Therefore, its low specificity has brought a lot of confusion to clinicians. For the diagnosis of ovarian cancer, tumor markers with higher specificity need to be discovered.

Method used

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  • Monoclonal antibody resisting ovarian cancer and application thereof
  • Monoclonal antibody resisting ovarian cancer and application thereof
  • Monoclonal antibody resisting ovarian cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation of embodiment 1 hybridoma

[0024] 1.1 Animal immunization Take 6-8 weeks old female BALB / c mice, each with 1-2×10 ovarian cancer cells 6 SK-OV-3 cells were immunized by intraperitoneal injection 4 times at intervals of 3 weeks. Blood was collected from the inner canthus of the mice before each immunization, and the antibody titer of the mouse serum was detected by indirect cell ELISA. When the serum antibody titer of the immunized mice reached the maximum and no longer increased, the mouse splenocytes were taken for fusion. 3 days before fusion Boost immunity 1 time.

[0025] 1.2 Indirect cell ELISA test Inoculate 2×10 cells on a 96-well plate 5 SK-OV-3 cells per well (Shanghai Baili Biotechnology Co., Ltd.), until the cell growth and confluence reached 80%, fixed with 95% ethanol, washed 3 times with PBS, permeabilized with 0.2% Triton-X-100 for 20 min, and then used 50g / LBSA was blocked at 37°C for 2 hours, 100 μL of immunized mouse serum of differ...

Embodiment 2

[0027] Example 2 Preparation and purification of monoclonal antibody ascites

[0028] Female BALB / c mice aged 8-10 weeks were injected intraperitoneally with 0.5 mL of paraffin oil, and 10 days later, they were injected intraperitoneally with well-growing hybridoma cells NM003-1 and NM003-2 at approximately 1×10 6 After 1-2 weeks, the ascites was aspirated, after 1 hour at 37°C, and overnight at 4°C, the ascites were centrifuged the next day and purified by ProteinG affinity chromatography to obtain purified monoclonal antibodies NJ003-1 and NJ003-2 .

Embodiment 3

[0029] Identification of embodiment 3 monoclonal antibody

[0030] 3.1 Identification of monoclonal antibody Ig subclass: the purified monoclonal antibody was diluted with PBS 1:10000, and operated according to the instructions of the assay kit. The subclasses of monoclonal antibodies NJ003-1 and NJ003-2 are IgG, and the light chain is κ chain.

[0031] 3.2 Determination of monoclonal antibody titer: Dilute the purified monoclonal antibodies NJ003-1 and NJ003-2 with PBS, respectively, take 100 μL and add them to 96-well plates coated with SK-OV-3 cells, and use indirect cell ELISA A 450 The titer is the maximum dilution of the monoclonal antibody that can immunoreact with the coated cells. The titer of monoclonal antibody NJ003-1 is 8×10 5 , the titer of monoclonal antibody NJ003-2 was 4×10 5 . The hybridoma cell line NM003-1 was deposited in China Center for Type Culture Collection on August 31, 2011, the deposit address is Wuhan, Wuhan University, and the deposit number...

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Abstract

The invention discloses a monoclonal antibody resisting the ovarian cancer and application thereof. A hybridoma cell strain NM003-1 secreting the monoclonal antibody resisting the human ovarian cancer is preserved in China Center for Type Culture Collection on September 26, 2011 with the preservation number of CCTCC NO: C201174. The hybridoma cell strain NM003-1 secretes the monoclonal antibody NJ003-1 resisting the human ovarian cancer. Expression of a specific antigen of the monoclonal antibody NJ003-1 in ovarian cancer tissue is higher than that in ovarian benign disease tissue; the positive expression rate of the antigen in a poorly differentiated patient is higher than that in a well differentiated patient. The monoclonal antibody NJ003-1 can effectively inhibit clone formation of ovarian carcinoma cells SK-OV-3 in soft agar and can remarkably inhibit growth of human ovarian cancer transplantation tumors in a nude mouse. Obviously, the monoclonal antibody is expected to become medicine for treating the ovarian cancer or to be prepared into a diagnostic reagent for detecting the ovarian cancer.

Description

technical field [0001] The invention belongs to the field of medical diagnosis, and relates to an anti-ovarian cancer monoclonal antibody and its application. Background technique [0002] Ovarian cancer is one of the most common malignant tumors of women's reproductive organs. Its incidence rate ranks third after cervical cancer and uterine body cancer, but its case fatality rate ranks first. In recent years, its incidence rate has been on the rise. A malignant disease that seriously threatens women's lives. Due to the anatomical position of the ovary deep in the pelvic cavity, the onset is hidden, and the early clinical symptoms and signs are not typical. About 70% of the patients are already in the advanced stage when they see a doctor, and most of them have metastasized, resulting in most patients losing the chance of radical surgery, leading to five The annual survival rate is low, less than 30%. [0003] Sugar chain antigen 125 (CA125) is a glycoprotein detected from...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/18G01N33/577G01N33/574A61K39/395A61P35/00
CPCC07K16/18
Inventor 潘世扬徐建王芳黄珮珺
Owner 潘世扬
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