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Tumor detection kit and its special substance for recognizing n-acetylglucosamine

A technology of acetamido and kits, applied in the direction of measuring devices, recombinant DNA technology, DNA / RNA fragments, etc., can solve problems such as difficult replication and unclear structure, and achieve the effect of improving early diagnosis rate

Inactive Publication Date: 2016-05-18
范飞舟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The exact structure of most current tumor markers is still unclear and difficult to reproduce

Method used

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  • Tumor detection kit and its special substance for recognizing n-acetylglucosamine
  • Tumor detection kit and its special substance for recognizing n-acetylglucosamine
  • Tumor detection kit and its special substance for recognizing n-acetylglucosamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1. Discovery and identification of N-acetylglucosamine for tumor detection

[0079] The inventors of the present invention have found that the content of N-acetylglucosamine in the blood of tumor patients is significantly different from that in healthy people.

[0080] In the present invention, the blood of tumor patients is directly analyzed by SELDI and MELDI mass spectrometry in vitro, to obtain the special sugar structure map of tumor patients, and to find the specific spectral lines of tumors, thereby discovering tumor-specific sugar structures, Glycoprotein, what we have discovered so far is the special sugar group of GlcNAc (N-acetylglucosamine, N-acetylglucosamine), and then the target sugar obtained by separation and purification by polyacrylamide electrophoresis, high pressure liquid chromatography, and lectin affinity chromatography protein.

[0081] We tried the method of affinity chromatography (Mu-GlcNAc was attached to the monoclonal antibody Ma6...

Embodiment 2

[0090] Example 2. A method for screening specific binding bodies to detect tumors using N-acetylglucosamine (sandwich method)

[0091] Determination of Mu-GlcNAc antigen in sandwich mode, the principle is as follows figure 2 shown.

[0092] (1) Materials and methods

[0093] 1 Reagents and tumor patient serum samples

[0094] 1.1 Reagent Ma695 monoclonal antibody (Mab), CanAgDiagnosticsAB, Sweden, biotin (biotin)-linked Aptamer (the aptamer shown in sequence 1 prepared as in Example 1, biotin-labeled), Mu-GlcNAc antigen, self-affinity and chromatographic extraction (see Example 1); it can also be purchased from Beijing Yonghan Xinggang Biotechnology Co., Ltd.

[0095] 1.2 Serum specimens of cancer patients The blood samples of cancer patients were obtained from the Cancer Hospital of the Chinese Academy of Medical Sciences and the 301 Hospital of the People's Liberation Army, and the diagnosis of the patients was confirmed by pathology.

[0096] 2 methods

[0097] 2.1 Pr...

Embodiment 3

[0116] Embodiment 3, single conjugate assay method detects tumor

[0117] The principle of the single-conjugate assay method is as follows: Figure 4A and Figure 4B , using the specific binding body aptamer (shown in sequence 1) of N-acetylglucosamine. Single-conjugate analysis uses only N-acetylglucosamine-specific binding body as the recognition body; single-conjugate analysis uses N-acetylglucosamine-specific binding body to directly identify N-acetylglucosamine in the detection object; The essence of conjugate analysis is the specific combination of conjugate and -GlcNAc.

[0118] The specific method is as follows:

[0119] (1) Materials and methods:

[0120] 1 Reagents and tumor patient serum samples

[0121] 1.1 Reagents: aptamer biotin-MA153-A, Beijing Yonghan Xinggang Biotechnology Co., Ltd.; MA153 antigen, Beijing Yonghan Xinggang Biotechnology Co., Ltd.; Streptavidin, SigmaUSA.

[0122] 1.2 Serum specimens of cancer patients The serum samples of cancer patient...

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Abstract

Disclosed are a kit for detecting tumours and an aptamer specifically binding to N-acetylglucosamine. The kit contains a substance for detecting tumours, and a substance specifically binding the substance for detecting tumours to N-acetylglucosamine.

Description

technical field [0001] The invention relates to a kit for detecting tumors and its special substance for recognizing N-acetylglucosamine. Background technique [0002] Since the 1980s, experts have used hybridoma technology to obtain tumor-specific macromolecular glycoprotein antigens (carbohydrateantigen, CA), and developed a monoclonal antibody recognition system. So far, in the field of in vitro early diagnosis reagents for tumors, there are two analysis methods: competitive analysis and non-competitive sandwich analysis. From the perspective of laboratory detection, tumor markers are divided into serum / plasma tumor markers and histiocytic tumor markers. Serum / plasma tumor markers are roughly divided into three categories: 1) Embryonic antigens. Antigens that are expressed in embryos, not expressed in normal adults, and re-expressed with tumorigenesis, such as AFP and CEA. 2) Tumor-associated glycolipids and glycoproteins. CA199, CA50, CA242, CA724, CA125, CA153, etc....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/574C12N15/115
CPCC12N15/115G01N33/574G01N33/577
Inventor 范振符
Owner 范飞舟
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