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Tumor marker CD25 autoantibody and application thereof

A tumor marker, tumor technology, applied in the field of immunology, can solve the problems of low sensitivity and poor specificity, and achieve the effect of high sensitivity and high specificity

Inactive Publication Date: 2013-05-29
尉军
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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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  • Tumor marker CD25 autoantibody and application thereof
  • Tumor marker CD25 autoantibody and application thereof
  • Tumor marker CD25 autoantibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The binding process of CD25 antigen polypeptide to serum and plasma IgG

[0038] It can be seen from Figure 1 that when the CD25 concentration is 5-10 μg / ml, the SBI value gradually decreases with the increase of the concentration, and when the CD25 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 CD25 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 CD25 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 specific binding between the antigen polypept...

Embodiment 2

[0040] kit preparation

[0041] 1 Reagents See Tab.2-9 for reagent preparation.

[0042]

[0043]

[0044] 2 operation

[0045] (1) Coating: Dilute the working antigen and reference antigen with coating solution to the working concentration, coat on the microtiter plate, and 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 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, add 100 μl of substrate color development solution to each well, and keep at room temperature in the dark for 15 to 30 mi...

Embodiment 3

[0051] Detection of CD25 Auto IgG Antibody in Patients with Lung Cancer

[0052] 1 Sample collection: 501 plasma samples from tumor patients and healthy people 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 consisted of 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 Detection results: From Tab.10-11, it can be known that the area under the ROC curve (AU) of the IgG antibody ROC curve (AU) of the CD25 antigen polypeptide in the plasma of lung cancer patients is 0.7, the sensitivity is 35%, and the specificity is 90%. The positive rate of IgG antibody binding to CD25 polypeptide antigen in plasma of patients with lung cancer was significantly higher than that of healthy group (Z=-7.48, P<0.001). The above data fully...

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Abstract

The invention discloses a tumor marker CD25 autoantibody and application of the tumor marker CD25 autoantibody, belonging to the technical field of immunology. The invention provides an amino acid sequence of antigen of immune response regulation gene CD25. The CD25 polypeptide antigen is used for detecting corresponding specific autoantibody in blood of lung cancer and esophageal cancer patients; and the autoantibody can be used as a tumor marker to evaluate risk level of occurrence of lung cancer and esophageal cancer. The antigen polypeptide and the antibody can be used for preparing early-stage tumor diagnostic reagent and developing target drugs for treating tumor.

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 autoantibodies, and there are both tumor antigens and autoantibodies against the tumor antigens in the serum of cancer patients. Therefore, both antibodies can be used to detect tumor antigens, and antigens can also be used to detect tumor autoantibodies, but the specificity and sensitivity of using tumor autoantibodies to detect tumors are much higher than using tumor antigens to detect tumors. 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 autoantibodies in normal peopl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28G01N33/68G01N33/574
Inventor 尉军孙世龙关松磊李光辉刘宝刚
Owner 尉军
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