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Application of peptidylarginine deiminase 1 (PAD1) to preparation of reagent for clinical diagnosis of tumors

A technology of peptidyl arginine deiminase and diagnostic reagent, which is applied in the application field of peptidyl arginine deiminase 1 in the preparation of tumor clinical diagnostic reagents, and can solve the problems of distribution in different tissues and the like

Active Publication Date: 2015-02-18
SHANDONG XINCHUANG BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PAD1 (peptidyl arginine deiminase 1, Peptidylarginine deiminase 1, or PADI1) and PAD4 are both peptidyl arginine deiminases, they have different tissue distributions. At present, there is no research on the role of PAD1 in tumors. Related reports on blood markers

Method used

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  • Application of peptidylarginine deiminase 1 (PAD1) to preparation of reagent for clinical diagnosis of tumors
  • Application of peptidylarginine deiminase 1 (PAD1) to preparation of reagent for clinical diagnosis of tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1  PAD1 test example 1

[0054] PAD1 was used as a tumor blood marker to detect the blood of tumor patients. The number of tumor patients and the types of tumors are shown in Table 1.

[0055] The detection method is:

[0056] (1) Dilute the blood sample to be tested 10 times with carbonate buffer (pH 9.6), add it to a blank microtiter plate, and incubate at 37°C for 2 hours;

[0057] (2) discard the liquid in the hole, and wash the plate three times with PBST washing solution (the PBST washing solution refers to the PBS solution plus Tween-20, the pH of the PBS solution is 7.4, and the concentration of Tween-20 is 0.1%, percent by volume) ;

[0058] (3) Add 200 μL / well of blocking solution (0.5% BSA, mass percent) to each well, and incubate at 37°C for 1 hour;

[0059] (4) Discard the liquid in the well, and wash the plate three times with PBST washing solution;

[0060] (5) Add 100 μL / well of PAD1 antibody dilution (1:2000 dilution) to each group, and incubate at 37°C for 1 hour;

[0061] (6) Di...

Embodiment 2  PAD1 test example 2

[0074] Using PAD1 as a tumor blood marker, the patient's blood was detected, and the detection method was the same as in Example 1. At the same time, with CEA, CA199, F / PSA, CA125, PSA, CA242, AFP, HCG, NSE in the prior art as tumor blood markers, the blood of tumor patients is detected (the detection method is a conventional method in the prior art). Method; the blood sample detection data of these tumor blood markers in the prior art is the patient outpatient data of a certain hospital, and this data can be used as one of the basis for diagnosing whether a patient has a tumor; the present invention uses the blood samples of some patients to detect PAD1 ). The positive rate of each tumor blood marker was compared, and the cross-comparison between PAD1 and each tumor marker.

[0075] Results: The number of patients (a total of 906 cases) and the number of positive cases are shown in Table 3. It can be seen from Table 3 that PAD1 can be used as a tumor blood marker with good ...

Embodiment 3  PAD1 test example 3

[0080] Using PAD1 as a blood marker, it is effective for hepatitis B, general inflammation (patients with increased white blood cells, neutrophils, and decreased lymphocyte ratio), kidney disease (including nephrotic syndrome, renal failure, nephritis), uremia (including uremia and urinary Road infection) patients' blood was tested, the number of patients and the type of disease are shown in Table 4.

[0081] The detection method is the same as in Example 1.

[0082] Result: The number of positive cases and the positive rate are shown in Table 4. As can be seen from Table 4, PAD1, as a blood marker, can also be used to detect hepatitis B, common inflammation (with white blood cells, neutrophils, symptoms of decreased lymphocyte ratio) ), kidney disease (including nephrotic syndrome, renal failure, nephritis), uremia (including uremia and urinary tract infection) and other diseases, the test results can be used as one of the basis for clinical diagnosis.

[0083] Table 4

[0...

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Abstract

The invention discloses an application of peptidylarginine deiminase 1 (PAD1) to preparation of a clinically diagnostic reagent or detection object for diagnosing tumors. The tumors are selected from uterine cancers, liver cancers, gastric cancers, esophageal cancers, ovarian cancers and lung cancers. When PAD1 is specially applied, an antibody against PAD1 is prepared by a conventional method, and a qualitative or quantitative method and a matching kit for detecting PAD1 are established. The invention also discloses a blood diagnosis reagent for diagnosing tumors. The blood diagnosis reagent comprises the PAD1 antibody, a horseradish peroxidase-immunoglobulin G (HRP-IgG) antibody and conventional reagents in the blood diagnosis reagent, wherein the conventional reagents in the blood diagnosis reagent include a carbonate buffer solution, a phosphate buffer solution-Tween (PBST) lotion, a blocking solution, developing solutions and a stop solution. PAD1 is found for the first time to be capable of being applied as a tumor blood marker. Experiments prove that PAD1 has feasibility as the tumor marker.

Description

technical field [0001] The invention relates to the application of peptidyl arginine deiminase 1 as a tumor blood marker in the preparation of tumor clinical blood diagnosis reagents. Background technique [0002] Peptidyl arginine deiminase (Peptidylarginine deiminase, PAD or PADI) is an enzyme in human tissue. At present, a total of five PAD enzymes (ie, PAD / PADI1, 2, 3, 4 and 6) have been found. These enzymes are encoded by a cluster of genes located on the lp36 region of the human chromosome and have distinct tissue distributions. It can perform post-translational modification on some other tissue proteins in the presence of calcium ions. This enzyme can catalyze the amino group of arginine (arginine) in the polypeptide chain into a carbonyl group, thereby converting arginine into citrulline (citrulline). Citrulline is an unnatural amino acid. This process of converting arginine to citrulline in polypeptides catalyzed by PAD is called citrullination. After citrullina...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/53
CPCG01N33/573G01N33/574
Inventor 常晓天邢艳秋马芳杨冬霞
Owner SHANDONG XINCHUANG BIOLOGICAL TECH CO LTD
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