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Amino acid sequence for detecting esophageal cancer marker P16 epitope and application of amino acid sequence

An epitope, esophageal cancer technology, applied in the field of immunology, can solve problems such as low expression rate, and achieve the effect of high specificity and high sensitivity

Pending Publication Date: 2021-10-26
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In domestic research on gastric adenocarcinoma, ovarian serous cystadenocarcinoma and endometrial cancer, it was found that the expression of P16 gene in cancer tissue showed an upward trend, but its expression rate was lower in the middle and late stage of cancer than in the early stage of cancer, which suggested that P16 It may become an effective indicator for distinguishing benign from malignant lesions, the severity of the disease, and judging the prognosis

Method used

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  • Amino acid sequence for detecting esophageal cancer marker P16 epitope and application of amino acid sequence
  • Amino acid sequence for detecting esophageal cancer marker P16 epitope and application of amino acid sequence
  • Amino acid sequence for detecting esophageal cancer marker P16 epitope and application of amino acid sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1P16

[0026] Process polypeptide antigen binding serum and plasma IgG embodiment according 1P16

[0027] Depend on figure 1 It found that, when the concentration of P16 5 ~ 10μg / ml, with increasing concentration, SBI value is gradually decreased, when the concentration of the antigen polypeptide P16 10 ~ 15μg / ml, with increasing concentration, SBI values ​​gradually increase. This SBI binding curves show that when the P16 antigen polypeptide is a lower concentration 5μg / ml (0.5μg / well), the plate bottom 96 well microtiter plate is not covered, resulting in a high non-specific reactions, so in this case the value of SBI high, false positive results; P16 antigen with the increase of concentration of polypeptide, the antigen gradually covered the entire bottom of the plate, which block the action of apparent non-specific reaction was gradually reduced to the lowest non-specific reaction 10μg ml / , this time P16 antigen-specific IgG antibody binding to the polypeptide appeared, and ...

Embodiment 2

[0028] Example 2 Kit Preparation

[0029] 1, the reagent is formulated as follows:

[0030] Antigenic package was buffer:

[0031] Sodium Sodium 0.1g

[0032] PBS (0.01M, pH 7.4) 100ml 4 ° C preservation

[0033] 0.1M PBST wash buffer (1000ml volume):

[0034] NaCl 8G

[0035] KCL0.2G

[0036] NA 2 HPO 4 .12h 2 O 2.9G

[0037] K h 2 PO 4 0.2g

[0038] TWEEN-20 0.5ml

[0039] Save 4 ° C, 10 times dilution before use

[0040] Sample dilution analysis solution:

[0041] BSA1.0G

[0042] 0.01MPBS UP to 100ml-20 ° C Save

[0043] Terminate buffer:

[0044] Concentrated H 2 SO 4 10.9ml

[0045] Distilled water 89.1ml

[0046] Up to 100ml room temperature Save

[0047] Work antigen:

[0048] P16 polypeptide antigen 5mg

[0049] 67% acetic acid 1ml

[0050] -20 ° C storage, dilute to 7.5 ~ 15μg / ml before use

[0051] Reference antigen:

[0052] GAG 5mg

[0053] 67% acetic acid 1ml

[0054] Dilute it to 20 μg / ml, -20 ° C preservation before use

[0055] Second anti-standard solution:

[...

Embodiment 3

[0067] Example 3 In particular, P16, IgG antibody detection, esophageal cancer

[0068] 1. Sample Source: Collect 400 patients with tumor and healthy plasma samples. 200 cases of health group, age, and average age of 52.9 ± 28.24 years. 200 cases of esophageal cancer, with an average age of 60.27 ± 8.33 years. The health group and esophageal cancer group have comparable (P> 0.05) in gender, age matching.

[0069] 2, the test results: It is known from Table 1 that the IgG antibody ROC curve of P16 antigen polypeptide in the plasma of esophageal cancer is 0.682, the sensitivity is 26.0%, and the specificity is 90%. figure 2 The ROC curve of esophageal cancer was detected for P16 antigen polypeptide. In the detection of IgG autoantibodies in patients with esophageal cancer, the expression of P16 autoantibody in the plasma in esophageal cancer has significant differences (P = 0.000) compared to the health control group (P = 0.000). In the installment statistics, there is a significant...

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PUM

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Abstract

The invention relates to an amino acid sequence for detecting esophageal cancer marker P16 epitope and application of the amino acid sequence, and belongs to the technical field of immunology. The invention provides an antigen amino acid sequence of a tumor cancer suppressor gene P16, a P16 polypeptide antigen is used 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, and the antigen polypeptide and an antibody thereof can be used for preparing an esophageal cancer early diagnosis reagent.

Description

Technical field [0001] The present invention belongs to the technical field of immunology, it is a preparation for the early diagnosis of esophageal cancer agent. Background technique [0002] Numerous studies show that the serum or plasma of tumor-associated antigens can induce an antigenic epitope, the presence of both the tumor antigen in the serum of cancer patients, there is an antigen epitope for the tumor. Thus, using both tumor antigen antibody may be detected using a tumor antigen epitope, but the use of tumor specificity and sensitivity of detection of tumor epitopes than the detection of tumor using the tumor antigen is much higher. Many tumor-associated antigen is present not only in cancer patients, but also in normal human body is present, the detection of tumor associated antigens as the basis for credible differential diagnosis. The epitope normal levels in the body can not detect very low or non-existent, if the body epitope levels were significantly higher, it i...

Claims

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Application Information

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IPC IPC(8): C07K14/47G01N33/574
CPCC07K14/4748G01N33/57407G01N33/57488
Inventor 刘林林皇甫明美
Owner JILIN UNIV
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