Establishment method of CD306 sandwich ELISA kit and application thereof

A technology for establishing methods and kits, which can be applied to measuring devices, instruments, and disease diagnosis, and can solve problems such as poor accuracy, lack of foresight, and high cost

Inactive Publication Date: 2016-04-20
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the diagnosis of colorectal cancer mainly includes colonoscopy, pathological biopsy, CEA detection, abdominal CT, etc. The patient's examination process is complicated, the cost is high, and the diagnosis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0015] The establishment of embodiment 1CD306 sandwich ELISA kit

[0016] (1) Use biosensors to identify the epitopes recognized by the anti-CD306 monoclonal antibody, and understand the spatial positional relationship of the epitopes recognized by each antibody

[0017] In step (1), acetic acid buffer is added to the sample reaction pool of the biosensor respectively, and then CD306-Fc fusion protein is added (the eukaryotic expression of CD306 fusion protein includes the following specific processes: using CD306cDNA as a template, amplifying the cDNA with primers , the cloned CD306 was inserted into the vector, a eukaryotic expression vector was constructed, a cell line was transfected, a transfected cell line that stably secreted the CD306-Fc fusion protein was cloned, the transfected cells were cultured and the fusion protein was purified) and human IgG standard ( (purchased from Gibco Company), and observe the binding amount of CD306-Fc and human IgG standard substance to...

Embodiment 2

[0021] Application of Example 2CD306 Sandwich ELISA Kit

[0022] (1) Use fluorescently labeled anti-CD305 monoclonal antibody to conduct early diagnosis of colorectal cancer by flow cytometry, and determine the quantitative standard for early diagnosis of colorectal cancer by flow cytometry

[0023] Using FITC (fluorescein isothiocyanate) to label anti-CD306 monoclonal antibody to prepare fluorescent antibody FITC-anti-CD306, isolate PBMC (peripheral blood mononuclear cells) from colorectal cancer patients for intracellular cytokine staining, and analyze the colorectal by flow cytometry Relationship between cancer stage and Lair expression level.

[0024] (2) Use the CD306 sandwich ELISA kit to detect the level of CD306 in the serum of patients with colorectal cancer, and use the standard to calculate the serum concentration of CD306 in the serum

[0025] The anticoagulant blood of patients with colorectal cancer was extracted, and the level of CD306 in the serum of patients ...

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Abstract

The invention provides an establishment method of a CD306 sandwich ELISA kit and its application. According to the establishment method, epitope of anti-CD306 monoclonal antibody is identified to establish the CD306 sandwich ELISA kit; and then, sensitivity of the kit is raised. The kit is used in evaluating the relationship between serum level of CD306 and prognosis of patients with colon and rectal cancers. Firstly, quantitative standard for early diagnosis of colon cancer by flow cytometry is determined; then, CD306 level in serum of patients with colon and rectal cancers is detected by the use of the CD306 sandwich ELISA kit; and finally, patients with colon cancer undergo long-term follow-up, detection indexes are recorded, and the relationship between serum level of CD306 and prognosis of patients with colon and rectal cancers is evaluated. The kit provided by the invention is economical and convenient, has good compliance, can be used to raise detection rate of early colon cancer and reduce death from early colon cancer.

Description

technical field [0001] The invention belongs to the technical field of early diagnosis of cancer, and in particular relates to the establishment method and application of a CD306 sandwich ELISA kit. Background technique [0002] Colorectal cancer is a common malignant tumor, and its incidence rate ranks third among malignant tumors, and ranks fifth among malignant tumors in my country. With the enhancement of people's health awareness and the improvement of medical diagnosis level, it is expected that the incidence of colorectal cancer in my country will continue to increase. Only 5% of colorectal cancers are diagnosed in the early stage, and 60% to 70% of colorectal cancer patients are found to be in stage II or advanced stage. The 5-year survival rate after surgery is about 50%, and the recurrence rate after surgery is as high as 30%, which seriously affects people. health and quality of life. Because there are many treatment methods for early colorectal cancer and the p...

Claims

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Application Information

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IPC IPC(8): G01N33/574
CPCG01N33/57419G01N33/57446G01N2800/52G01N2800/54
Inventor 康振华王磊袁春华张文梁张剑涛岳勐
Owner JILIN UNIV
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