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Detection kit of galectin-3 binding protein and method and application thereof

A technology of galectin and detection kit, which is applied in the field of medical immunology and in vitro diagnosis, can solve the problems of low detection sensitivity and specificity, easy missed diagnosis of early needle biopsy, and difficulty in finding early patients, etc., and achieves detection cost Low, high sensitivity, good stability

Inactive Publication Date: 2017-05-31
广州赛太特生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have the following disadvantages: detection sensitivity and specificity are not high, it is difficult to find early patients; early needle biopsy is easy to miss diagnosis; invasive, difficult to be generally accepted by the public; expensive
But so far, there is no relevant report on the application of G3BP protein as a serum marker in the early screening or diagnosis of lung cancer

Method used

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  • Detection kit of galectin-3 binding protein and method and application thereof
  • Detection kit of galectin-3 binding protein and method and application thereof
  • Detection kit of galectin-3 binding protein and method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Select mouse anti-G3BP monoclonal antibody and rabbit anti-G3BP polyclonal antibody to carry out concentration gradient test, and determine the optimal working concentration of G3BP monoclonal antibody is 4 μg / mL, and the optimal working concentration of G3BP polyclonal antibody is 2.5 μg / mL.

[0048] 2. Preparation of microwell plates coated with G3BP monoclonal antibody

[0049] Dilute the mouse anti-G3BP monoclonal antibody to 4 μg / mL with coating buffer, and then use it to coat 96 microwell plates, 50 μL per well, and let stand at 37°C for 2 hours; then, discard the supernatant, and add 50 μL of After blocking with 10% goat serum for 1 hour, remove the liquid in the wells, and pat dry on clean absorbent paper; then add 350 μL of washing solution to each well, let it stand for 5-10 seconds, discard it, repeat 5 times, and finally pat on clean absorbent paper Dry.

[0050] 3. Preparation of standard solution

[0051] Dilute the recombinant G3BP with diluent to th...

Embodiment 2

[0069] Fasting venous blood was collected from lung cancer stage Ia patients (9 cases), lung cancer stage Ib patients (12 cases), lung cancer stage IIa patients (8 cases) and healthy volunteers (26 cases, age-matched), and the blood samples were placed at room temperature for 1- 2h, then centrifuged at 3000rpm for 10min, and then the separated serum was stored in -20°C refrigerator. Among them, the patients and healthy volunteers were inpatients and healthy people from the First Affiliated Hospital of Guangzhou Medical University and the Second Affiliated Hospital of Guangzhou Medical University, respectively. The lung cancer patients were diagnosed by pathological diagnosis and immunohistochemistry.

[0070] First, the kit was taken out of the refrigerator at 4°C, and placed at room temperature to allow the temperature of each serum sample to equilibrate to room temperature.

[0071] Next, take the microwell plate coated with G3BP monoclonal antibody, fix it on the frame, set...

Embodiment 3

[0079] Using the technical means and detection specifications commonly used by those skilled in the art, the detection kit prepared in Example 1 and the commercially available enzyme-linked immunosorbent assay kit were compared in terms of sensitivity, precision and stability, and the results were as follows:

[0080] 1. Sensitivity investigation

[0081] The detection range of the G3BP chemiluminescence kit of the present invention is 5-3000 pg / mL, and its detection sensitivity is 4.78 pg / mL (average concentration of blank wells + 3 times standard deviation). When using a commercially available enzyme-linked immunosorbent assay kit to detect G3BP in the patient's serum, the detection sensitivity of the kit is 42.5 pg / mL (the mean value of the concentration of the blank well + 3 times the standard deviation). Therefore, the sensitivity of the chemiluminescence kit for detecting G3BP in the present invention is much higher than that of the traditional enzyme-linked immunosorben...

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Abstract

The invention belongs to the technical field of medical immunity and in vitro diagnosis, and particularly relates to a detection kit of a galectin-3 binding protein and a method and application thereof. The detection kit provided by the invention is high in sensitivity, high in specificity, and capable of rapidly and qualitatively detecting the content of G3BP in human serum, capable of nondestructive sampling, easy to accept by the population, low in detection cost, simple operation procedures and good in stability, and has an important significance and real application value for the early screening and diagnosis and timely treatment of lung cancer.

Description

technical field [0001] The invention belongs to the technical field of medical immunology and in vitro diagnosis, and in particular relates to a detection kit for galectin-3 binding protein and its method and application. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality in the world, and the age of onset is 45-80 years old. In my country, about 400,000 patients die of lung cancer every year, and the number of cases is increasing year by year, seriously endangering public health. Lung cancer in the early stage is mostly asymptomatic or very mild, which is not enough to attract the attention of patients and misses the opportunity to diagnose early lung cancer. When patients come to the hospital with symptoms of cough, chest pain, and hemoptysis, 80% of them are already in the late stage. After surgery and comprehensive treatment, the 5-year survival rate of early lung cancer can reach 60%-90%; while most patients with ad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/57423G01N33/57484G01N33/6893
Inventor 汤郡葛晨晨陈颖
Owner 广州赛太特生物医学科技有限公司
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