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Application of group of tumor-associated antigens in preparation of kit for early screening of liver cancer

A tumor-associated antigen and kit technology, applied in the fields of molecular biology and oncology, to achieve good application prospects, good detection sensitivity and specificity, and the effect of reducing mortality

Pending Publication Date: 2021-06-01
PEOPLES HOSPITAL OF HENAN PROV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of six tumor-associated antigens, ALKBH1, PPP2R1A, BCAS4, ASTE1, UGT3A2, and FAM5C, to detect the expression of corresponding autoantibodies in the serum of patients with liver cancer has not been reported.

Method used

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  • Application of group of tumor-associated antigens in preparation of kit for early screening of liver cancer
  • Application of group of tumor-associated antigens in preparation of kit for early screening of liver cancer
  • Application of group of tumor-associated antigens in preparation of kit for early screening of liver cancer

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Experimental program
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Effect test

Embodiment 1

[0028] Example 1 Preparation of tumor-associated antigens

[0029] By using a prokaryotic expression system, six tumor-associated antigens, ALKBH1, PPP2R1A, BCAS4, ASTE1, UGT3A2, and FAM5C, were prokaryotically expressed and purified to prepare for the preparation of the kit.

[0030] The preparation process of the above six tumor-associated antigens is as follows:

[0031] (1) Using gene cloning technology to construct recombinant prokaryotic expression plasmids of six tumor-associated antigen proteins, ALKBH1, PPP2R1A, BCAS4, ASTE1, UGT3A2, and FAM5C;

[0032] (2) Expressing the target protein: Transform the constructed recombinant prokaryotic expression plasmids into Escherichia coli BL21(DE3), and use IPTG (isopropylthiogalactopyranoside) to induce the expression of the target protein;

[0033] (3) Purify the target protein: according to the tag carried by the target protein, use the traditional corresponding purification scheme to purify the target protein;

[0034] (4)...

Embodiment 2

[0036] The preparation of embodiment 2 kit

[0037] According to the principle of indirect ELISA, the invention prepares an autoantibody joint detection ELISA kit for screening and diagnosing early liver cancer. The principle of indirect enzyme-linked immunoassay is to connect the antigen to the solid-phase carrier, and the antibody to be tested in the serum sample is combined with it to form a solid-phase antigen-test antibody complex, and then the enzyme-labeled secondary antibody and the solid-phase antigen-test antibody The antibody in the antibody complex is combined to form a solid-phase antigen-test antibody-enzyme-labeled secondary antibody complex, and then measure the degree of color development after adding the substrate to determine the content of the antibody to be tested. The above-mentioned detection principle is as follows: figure 2 shown.

[0038] The preparation process of the ELISA kit for detecting early liver cancer of the present invention is as follows: ...

Embodiment 3

[0075] The usage method of embodiment 3 kit

[0076] The concrete using method of kit of the present invention is as follows:

[0077] 1. Incubation of Serum Samples

[0078] Dilute the serum sample to be tested with the sample diluent at a ratio of 1:100, then add the diluted serum sample to the reaction wells of the antigen-coated 48-well microplate plate, the sample volume is 100 μL / well, set Incubate for 1 h in a constant temperature incubator at 37°C, then remove the liquid in the reaction wells, and wash 5 times with washing solution, 3 min each time.

[0079] 2. Incubation of the enzyme-labeled secondary antibody

[0080] Dilute the horseradish peroxidase-labeled RecA protein with the secondary antibody diluent at a ratio of 1:80000, and then add the diluted horseradish peroxidase-labeled RecA protein to the reaction wells of the 48-well microtiter plate In this method, the sample volume was 100 μL / well, placed in a 37°C constant temperature incubator and incubated f...

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Abstract

The invention belongs to the field of molecular biology and oncology, and particularly relates to application of a group of tumor-associated antigens in preparation of a kit for early screening of liver cancer. The tumor-associated antigens for preparing the kit for early screening of liver cancer are combinations of ALKBH1, PPP2R1A, BCAS4, ASTE1, UGT3A2 and FAM5C. The invention provides the ELISA kit for early screening of liver cancer. The kit comprises a solid phase carrier and a tumor-associated antigen coated on the solid phase carrier, and the tumor-associated antigen is composed of ALKBH1, PPP2R1A, BCAS4, ASTE1, UGT3A2 and FAM5C. According to the invention, the six tumor-associated antigens are used as a combination for screening early liver cancer for the first time, and the kit has high detection sensitivity and specificity and is suitable for rapid screening of large-scale high-risk groups.

Description

technical field [0001] The invention belongs to the fields of molecular biology and oncology, and specifically relates to the application of a group of tumor-associated antigens in the preparation of a kit for early screening of liver cancer. Background technique [0002] Liver cancer is a malignant tumor of the liver, including primary liver cancer and secondary liver cancer; among them, primary liver cancer originates from the epithelial or mesenchymal tissue of the liver, and tumors originating from the epithelial tissue of the liver are the high incidence of liver cancer in my country. The molecular mechanism is not completely clear, and its pathogenesis is a multi-factor and multi-step complex process, which is affected by the dual factors of environment and diet. Epidemiological and experimental studies have shown that hepatitis B virus, hepatitis C virus, aflatoxin, drinking water pollution, alcohol, liver cirrhosis, sex hormones, nitrosamine compounds, trace elements,...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/57438G01N33/543G01N33/57488Y02A50/30
Inventor 许芳芳刘小转申丽君葛菲时明月
Owner PEOPLES HOSPITAL OF HENAN PROV
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