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Amino acid sequence for detecting tumor marker P16 antigenic epitope and application of amino acid sequence

A tumor marker, amino acid technology, applied in the field of immunology, can solve the problem of low expression rate and achieve the effect of high specificity and high sensitivity

Inactive Publication Date: 2014-11-26
尉军
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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 tumor marker P16 antigenic epitope and application of amino acid sequence
  • Amino acid sequence for detecting tumor marker P16 antigenic epitope and application of amino acid sequence
  • Amino acid sequence for detecting tumor marker P16 antigenic epitope and application of amino acid sequence

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] P16 The process of antigen peptide binding to serum and plasma IgG

[0034] Depend on figure 1 It can be seen that when the P16 concentration is 5-10 μg / ml, the SBI value gradually decreases with the increase of the concentration, and when the P16 antigen polypeptide concentration is 10-15 μg / ml, the SBI value gradually increases with the increase of the concentration. This SBI binding curve shows that when the P16 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 the concentration of the P16 antigen peptide is too high, it is a false positive result; as the concentration of the P16 antigen polypeptide increases, the antigen gradually covers the entire bottom of the plate, its blocking effect appears, and the non-specific reaction gradually decreases, and the non-specific reaction is the lowest at 10 μg / ml. The s...

Embodiment 2

[0036] kit preparation

[0037] Tab.2 Antigen coating buffer Sodium azide 0.1g PBS (0.01M, pH7.4) 100ml Store at 4°C

[0038]

[0039]

[0040]

[0041]

[0042]

[0043] 2 operations

[0044] (1) Coating: the working antigen and reference antigen are diluted to the working concentration with coating solution, and coated on the microtiter plate,

[0045] overnight at 4°C.

[0046] (2) Add plasma (primary antibody): Wash the ELISA plate 3 times with washing buffer, dilute the plasma to an appropriate concentration with the analysis solution, generally 1:200-1:500, 100 μl per well, incubate at 25°C or room temperature 2~3h;

[0047] (3) Secondary antibody incubation: wash with washing buffer for 3 to 5 times, dilute the secondary antibody standard solution IgG with the analytical solution, add 200 μl to each well, and incubate at 25°C / room temperature for 2 hours;

[0048] (4) Color development: wash with washing buffer 3 to 5 times...

Embodiment 3

[0051] lung cancer patients P16 Auto IgG antibody detection

[0052] 1 sample collection: 501 plasma samples from tumor patients and healthy individuals were collected. The healthy group consisted of 227 cases, with an average age of 57.07±10.36 years, including 134 males and 92 females. The lung cancer group included 274 cases, with an average age of 57.5±9.2 years, including 177 males and 97 females. The healthy group and the lung cancer group were matched in gender and age and were comparable ( P >0.05)

[0053] 2 test results : According to Tab.10-11, the area under the ROC curve (AU) of the IgG antibody ROC curve of the P16 antigen polypeptide in the plasma of lung cancer patients was 0.562, the sensitivity was 19.6%, and the specificity was 90.4%. The positive rate of IgG antibody combined with P16 polypeptide antigen in the plasma of patients with lung cancer was significantly higher than that of the healthy group ( Z = -2.269, P <0.05). The above data ful...

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Abstract

The invention discloses an amino acid sequence for detecting a tumor marker P16 antigenic epitope and application of the amino acid sequence, and belongs to the technical field of immunology. The invention provides an antigenic amino acid sequence of a tumor anti-cancer gene P16. The P16 polypeptide antigen is used for detecting corresponding specific antigenic epitope in the blood of a patient with lung cancer and esophagus cancer; the antigenic epitope can be used as a tumor marker for estimating the degree of risk for occurring the lung cancer and the esophagus cancer. And the antigenic polypeptide and an antibody thereof can be used for preparing tumor early-diagnosis reagent and developing a targeted drug for treating the tumors.

Description

technical field [0001] The invention belongs to the technical field of immunology, and relates to the preparation of tumor early diagnosis reagents and the development of targeted drugs for treating tumors. 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 all....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/30
Inventor 孙世龙尉军李光辉
Owner 尉军
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