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Application of esophageal cancer related antigen protein combination or specific antibody thereof in esophageal cancer detection kit

A technology for detecting kits and tumor-associated antigens, applied in the fields of molecular biology and oncology, to achieve the effects of screening and early detection, high detection accuracy, high sensitivity and specificity

Active Publication Date: 2020-06-23
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The latest genomics and proteomics analysis data of our research group, combined detection of 8 kinds of TAA, as early screening and diagnosis of esophageal cancer, there is no relevant literature report

Method used

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  • Application of esophageal cancer related antigen protein combination or specific antibody thereof in esophageal cancer detection kit
  • Application of esophageal cancer related antigen protein combination or specific antibody thereof in esophageal cancer detection kit
  • Application of esophageal cancer related antigen protein combination or specific antibody thereof in esophageal cancer detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Application of 8 kinds of tumor-associated antigens in the preparation of early screening esophageal cancer detection kit

[0038] According to the principle of indirect enzyme-linked immunosorbent method, the present application prepares an autoantibody combined detection ELISA kit that can be used for screening and diagnosing early esophageal cancer. The principle of indirect enzyme-linked immunoassay figure 2 As shown, the antigen is connected to the solid-phase carrier, and the antibody to be tested in the sample is combined with it to form a solid-phase antigen-test antibody complex, and then the enzyme-labeled secondary antibody and the antibody in the solid-phase antigen-test antibody complex Combine 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.

[0039] The ELISA kit used in this e...

Embodiment 2

[0040] The preparation of embodiment 2 ELISA kit

[0041] 1. Experimental materials and reagents:

[0042] (1) Eight tumor-associated antigen proteins (RECK, RFT2, PTCH1, NOTCH1, FAT4, AJUBA, P53 and YAP1);

[0043] (2) 96-well ELISA plate: 3590 (costar. USA);

[0044] (3) Coating solution: 50mM carbonate buffer solution, pH=9.6;

[0045] (4) Blocking solution: PBST buffer containing 2% (W / V) BSA;

[0046] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0047] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0048] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (Invitrogen);

[0049] (8) Washing solution: 0.01M PBST (phosphate) buffer at pH 7.4;

[0050] (9) Positive control serum: P53 positive control serum, that is, the serum of patients with esophageal cancer who were positive for P53 antibody by indirect ELISA and Western blot;

[0051] (10) Negative control serum: P53 negative control serum, that ...

Embodiment 3

[0072] The application process of embodiment 3 ELISA kit

[0073] 1. Sample source

[0074] 400 serum samples were collected from the State Key Laboratory of Esophageal Cancer Prevention and Control jointly established by the province and ministry, including 200 serum samples from normal people (control group) and 200 serum samples from patients with early esophageal cancer (early esophageal cancer group). 200 cases of normal human serum were collected from the healthy checkup center of the laboratory's cooperative hospital, without any tumor-related evidence. Among the 200 normal subjects, there were 100 males and 100 females, the age range was 40-70 years old, and the average age was 58.9±5.3 years old. 200 serum samples from patients with early esophageal cancer were obtained from patients with early stage (stage 0+stage I) esophageal cancer confirmed by histopathology, and none of them received radiotherapy or chemotherapy. Among the 200 patients with esophageal cancer, ...

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Abstract

The invention belongs to the fields of molecular biology and oncology, and specifically discloses application of tumor-associated antigens RECK protein, RFT2 protein, PTCH1 protein, NOTCH1 protein, FAT4 protein, AJUBA protein, P53 protein and YAP1 protein in preparation of an esophageal cancer detection kit. Especially, the protein is applied to preparation of a kit for early screening esophagealcancer detection, so that the detection efficiency and diagnosis efficiency of clinical esophageal cancer are greatly improved, and the protein is beneficial to wide popularization and application incommon laboratories. When the eight proteins are applied to an ELISA kit, the expression levels of eight TAA antibodies in a serum sample can be detected at the same time, the detection success rate is high, and the technical reproducibility is good. Correspondingly, the molecule capable of specifically detecting the expression level of the combination of RECK, RFT2, PTCH1, NOTCH1, FAT4, AJUBA, P53 and YAP1 can also be used as an esophageal cancer detection test reagent, is applied to an esophageal cancer detection kit, and also has relatively high specificity and sensitivity.

Description

technical field [0001] The invention belongs to the fields of molecular biology and oncology, and in particular relates to the application of esophageal cancer-related antigen protein combinations or specific antibodies thereof in esophageal cancer detection kits. Background technique [0002] Esophageal cancer (EC) is a malignant tumor originating from the epithelium of the esophageal mucosa. The pathological types include squamous cell carcinoma and adenocarcinoma. It is one of the common malignant tumors of the digestive tract. In 2018, the global incidence of esophageal cancer ranked seventh among malignant tumors (6.3 / 100,000), and its mortality rate ranked sixth (5.5 / 100,000). In the same year, the incidence rate (13.9 / 100000) and mortality rate (12.7 / 100000) of esophageal cancer in my country ranked fifth and fourth among malignant tumors, and new cases and deaths accounted for 53.7% and 55.7% of the global total respectively. . my country is the country with the hig...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/543G01N33/57407G01N33/57488
Inventor 王立东杨苗苗赵学科宋昕张毅周胜理范宗民王苒李贝韩雪娜
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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