Tumor-associated autoantibody and tumor marker combined detection kit

A technology of tumor markers and autoantibodies, which is applied in the field of combined detection kits for tumor-related autoantibodies and tumor markers, can solve the problems of patients missing early detection and treatment, tumor marker levels not significantly increased, and low sensitivity

Pending Publication Date: 2019-10-15
GENE TECH SHANGHAI COMPANY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the fact that patients in the early stage of cancer have no obvious discomfort, the tumor volume is small, the cancer site is hidden, and the level of tumor markers in some patients does not increase significantly, the current screening methods mainly based on imaging methods and specific tumor markers have low sensitivity , there are also problems of high false positives and insufficient accuracy, which will not only make some patients miss the opportunity of early detection and treatment, but also may increase the workload and economic burden of follow-up monitoring

Method used

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  • Tumor-associated autoantibody and tumor marker combined detection kit
  • Tumor-associated autoantibody and tumor marker combined detection kit
  • Tumor-associated autoantibody and tumor marker combined detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0085] Example 1. Tumor-associated autoantibodies corresponding to antigenic proteins and anti-tumor marker antibodies were coated with magnetic particles

[0086] The method of directly immobilizing antigenic proteins to magnetic particles is as follows: the recombinant tumor-associated autoantibodies corresponding to antigenic proteins P53, SOX2, CK8, HuD, CAGE, NY-ESO-1, GBU4- 5. P62, KRAS and MAGE A4. Add 2.5 μg recombinant protein to 200 μL magnetic particle suspension (0.05M MES, pH 5.0, containing 1.25×10 6 magnetic particles) and incubate for 2 hours at room temperature in the dark. Then 20 μL of 1 mg / mL EDC was added to the mixture and incubated overnight at room temperature in the dark. Wash 4 times with 1 mL of PBS (containing 1% BSA, 0.2% Tween-20 and 0.05% sodium azide, pH 7.4), 500 μL of blocking / preservation buffer (PBS-TBN containing 0.1% BSA, 0.02% Tween-20 and 0.05 % sodium azide, pH 7.4) to resuspend the magnetic particles and store in the dark at 2-8°C. ...

Embodiment 2

[0089] Example 2. Preparation of kits for detecting tumor-associated autoantibodies and tumor markers

[0090] According to the principles of indirect ELISA and double-antibody sandwich method, the present invention prepares a combined detection kit of multiple autoantibodies and tumor markers that can be used for early screening and diagnosis of lung cancer. The principle of indirect enzyme-linked immunoassay is to connect the antigen protein corresponding to the tumor-associated autoantibody to a solid-phase carrier, and the autoantibody to be tested in the sample is combined with it to form a solid-phase antigen-tested autoantibody complex, and then the enzyme-labeled anti-antibody The human IgG antibody combines with the autoantibody in the solid-phase antigen-test autoantibody complex to form a solid-phase antigen-test autoantibody-enzyme-labeled anti-human IgG antibody complex, and then measure the luminous intensity after adding a luminescent substance to determine Auto...

Embodiment 3

[0108] Embodiment 3, detection method

[0109] 1. Sample and Standard Preparation

[0110] Dilute humanized Anti-His antibody standard 1 twice to obtain standard 2, and then serially dilute to obtain standard 3-6;

[0111] Dilute CEA standard 1, NSE standard 1, and SCC standard 1 by 5 times to obtain CEA standard 2, NSE standard 2, and SCC standard 2, and then serially dilute to obtain CEA standard 3-6, NSE standard 3 ~6 and SCC standard 3~6;

[0112] Part of the serum was diluted 100 times with sample diluent for detection of autoantibodies.

[0113] 2. Detection of autoantibodies

[0114] 2.1 Make a standard curve: draw a standard curve with the luminous intensity as the Y axis and the logarithm of the antibody concentration as the base 10 as the X axis, and calculate the standard curve regression equation.

[0115] 2.1.1 Add 50 μL of standard substance 1-6 into 12 reaction cups respectively, and divide into 6 groups;

[0116] 2.1.2 Add 20 μL Vector-His magnetic particl...

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Abstract

The invention relates to a combined detection method based on a tumor-associated autoantibody and a tumor marker, a reagent and a kit. A combination of three or more tumor-associated autoantibodies corresponding to antigen protein and one or more tumor marker antibodies is adopted to optimize and design a detection reagent and the kit, sensitivity and accuracy of tumor diagnosis are greatly increased, and the method, the reagent and the kit are especially suitable for early diagnosis of a tumor.

Description

technical field [0001] The invention belongs to the technical field of diagnosis or detection of tumor-related markers, in particular, the invention relates to a combined detection kit of tumor-related autoantibodies and tumor markers. Background technique [0002] According to the "Global Cancer Statistics 2018" released by the World Cancer Research Foundation, it is estimated that the total number of new cancer cases will reach 18 million in 2018, and there will be 9.6 million cancer deaths worldwide, of which, the number of new cases in China will account for 3.804 million cases, The number of deaths accounted for 2.296 million cases. [0003] Although with the continuous advent of new drugs, new treatments, and new technologies, and the country’s continuous increase in investment in the medical field, the 5-year survival rate of cancer has increased from 30.9% in 2003-2005 to 37.2% in 2012-2015 However, due to the increase in the incidence rate, malignant tumors are sti...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N21/76G01N21/64
CPCG01N33/57484G01N33/57488G01N21/76G01N21/6428G01N21/6486
Inventor 刘浩李宾王东胡欣张涛
Owner GENE TECH SHANGHAI COMPANY
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