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Antigen epitope amino acid sequence for detecting esophageal cancer marker Survivin and application of amino acid sequence

An antigen epitope and esophageal cancer technology, applied in the field of immunology, can solve the problems of poor specificity and low sensitivity, and achieve high specificity

Pending Publication Date: 2021-09-24
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported antibody detection methods have low sensitivity and poor specificity, and the false negative rate can be as high as more than 50%.

Method used

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  • Antigen epitope amino acid sequence for detecting esophageal cancer marker Survivin and application of amino acid sequence
  • Antigen epitope amino acid sequence for detecting esophageal cancer marker Survivin and application of amino acid sequence
  • Antigen epitope amino acid sequence for detecting esophageal cancer marker Survivin and application of amino acid sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 The process of Survivin antigen polypeptide binding to serum and plasma IgG

[0026]When the concentration of Survivin was 5-10 μg / ml, the SBI value gradually decreased with the increase of the concentration. When the concentration of Survivin antigen peptide was 10-15 μg / ml, the SBI value gradually increased with the increase of the concentration. This SBI binding curve shows that when the Survivin antigen polypeptide is at a lower concentration of 5 μg / ml (0.5 μg / well), the bottom of the 96-well microtiter plate is not covered, resulting in high non-specific reactions, so the SBI value at this time If it is too high, it is a false positive result; as the concentration of Survivin antigen peptide increases, the antigen gradually covers the entire bottom of the plate, its blocking effect appears, and the non-specific reaction gradually decreases. The specific binding between the antigen polypeptide and the IgG antibody began to appear, and gradually increased ...

Embodiment 2

[0027] Embodiment 2 kit preparation

[0028] 1. Reagent preparation:

[0029] Antigen coating buffer:

[0030] Sodium azide 0.1g

[0031] PBS (0.01M, pH7.4) 100ml Store at 4°C

[0032] 0.1M PBST wash buffer (1000ml volume):

[0033] NaCl 8g

[0034] KCl0.2g

[0035] Na 2 HPO 4 .12H 2 O 2.9g

[0036] K H 2 PO 4 0.2g

[0037] Tween-20 0.5ml

[0038] Store at 4°C and dilute 10 times before use

[0039] Sample dilution analysis solution:

[0040] BSA 1.0g

[0041] 0.01MPBS up to 100ml-20℃

[0042] Stop buffer:

[0043] Concentrated H 2 SO 4 10.9ml

[0044] Distilled water 89.1ml

[0045] up to 100ml store at room temperature

[0046] Working antigen:

[0047] Survivin Polypeptide Antigen 5mg

[0048] 67% acetic acid 1ml

[0049] Store at -20°C, dilute to 7.5-15 μg / ml with coating solution before use

[0050] Reference antigen:

[0051] gAg 5mg

[0052] 67% acetic acid 1ml

[0053] Dilute to 20μg / ml with coating solution before use, store at -20°C

...

Embodiment 3

[0066] Example 3 Detection of Survivin's own IgG antibody in patients with esophageal cancer

[0067] 1. Sample source: 400 plasma samples from tumor patients and healthy people were collected. Healthy group of 200 cases, age, mean age was 52.9 ± 28.24 years old. There were 200 patients in the esophageal cancer group, with an average age of 60.27±8.33 years. The healthy group and the esophageal cancer group were matched in gender and age and were comparable (P>0.05).

[0068] 2. Test results: It can be seen from Table 1 that the area under the ROC curve (AUC) of the IgG antibody to Survivin antigen polypeptide in the plasma of patients with esophageal cancer was 0.611, the sensitivity was 15.0%, and the specificity was 90%. figure 1 ROC curve for the detection of esophageal cancer by Survivin antigen peptide. In the detection of IgG autoantibodies in patients with esophageal cancer, the expression of Survivin autoantibodies in the plasma of patients with esophageal cancer w...

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Abstract

The invention relates to an antigen epitope amino acid sequence for detecting an esophageal cancer marker Survivin and application of the antigen epitope amino acid sequence, and belongs to the technical field of immunology. The invention provides the antigen amino acid sequence of a tumor cancer suppressor gene Survivin, the Survivin polypeptide antigen is applied for detecting a corresponding specific antigen epitope in the blood of an esophageal cancer patient, the antigen epitope can be used as a tumor marker for evaluating the risk degree of esophageal cancer, the antigen polypeptide and an antibody of the antigen polypeptide can be used for preparing an esophageal cancer early diagnosis reagent, and reliable data is provided for research of new tumor drugs.

Description

technical field [0001] The invention belongs to the technical field of immunology, in particular to an amino acid sequence and application of Survivin antigen epitope for detection of esophageal cancer marker. Background technique [0002] A large number of studies have shown that tumor-associated antigens in serum or plasma can induce the body to produce antigenic epitopes. In the serum of cancer patients, there are both tumor antigens and antigenic epitopes for the tumor antigens. Therefore, both antibodies and antigens can be used to detect tumor antigens, but the specificity and sensitivity of tumor antigen epitopes to detect tumors are much higher than those of tumor antigens. Many tumor-associated antigens exist not only in tumor patients, but also in normal people, so the detection of tumor-associated antigens is not reliable as a basis for diagnosis. However, the content of antigenic epitopes in normal people is very low and cannot be detected or does not exist at a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47G01N33/574G01N33/68
CPCC07K14/4703G01N33/57488G01N33/57407G01N33/6893G01N2333/4704
Inventor 刘林林皇甫明美
Owner JILIN UNIV
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