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Liver cancer biomarker and kit for detecting liver cancer

A technology of biomarkers and detection reagents is applied in the field of liver cancer biomarkers and kits for detecting liver cancer to achieve the effect of specific diagnosis

Active Publication Date: 2022-03-11
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sialic acid is an important part of the modification of cell membrane glycoproteins, but so far there is no relevant research report on the use of sialic acid (Neu5Gc) modified IgG in serum as a diagnostic marker for liver cancer

Method used

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  • Liver cancer biomarker and kit for detecting liver cancer
  • Liver cancer biomarker and kit for detecting liver cancer
  • Liver cancer biomarker and kit for detecting liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The IgG polyclonal antibody involved in the present invention is prepared by a laboratory, and the separation and purification steps of the mouse IgG polyclonal antibody are as follows:

[0027] (1) Dilute mouse serum in PBS at a ratio of 1:10, filter through a 0.22 μm filter membrane, and incubate with a Protein G agarose column at 4°C for 6 hours;

[0028] (2) Wash the purification equipment with about 30 mL of 20% aqueous ethanol;

[0029] (3) Use Binding Buffer (0.1M NaCl, 20mM NaHPO 4 , pH=7.0-7.4) balance the purification equipment, after the purification equipment indication is stable, adjust the indication to "0";

[0030] (4) Sample loading: load the sample obtained in step (1) at a speed of 1mL / min (injection speed adjusted to 12-18), after the serum premixed liquid in the column is loaded The effluent can be received by rising rapidly, that is, "flow through"), and the Binding Buffer is used to balance the purification equipment;

[0031] (5) Equilibrium: ...

Embodiment 2

[0037] The preparation of the lamprey immune protein LIP used in this example refers to Patent No. 201310501366.2, a Chinese invention patent named "Lamprey Li protein, preparation method and application in the preparation of drugs for the prevention and treatment of tumor diseases". The lamprey immune protein LIP can be obtained by isolating from the culture medium of lamprey Lie bodies, or obtained through a heterologous expression system, such as an Escherichia coli prokaryotic expression system, preferably a pColdI expression vector, to efficiently obtain the recombinant protein rLiprotein.

[0038] The preparation of the isolation of lamprey immune protein LIP from the culture medium of lamprey Listeri bodies was carried out as follows:

[0039] a. Take fresh Lamprey Lee's bodies and put them into trypsin for overnight digestion at 4°C;

[0040] b. Collect digested cells and wash twice with PBS;

[0041] c. Transfer to serum-free 1640 culture medium and cultivate for 72 ...

Embodiment 3

[0052] The invention provides a test kit for detecting liver cancer, which includes a coating reagent, a washing solution, a capture reagent, a detection reagent, a chromogenic reagent and a termination reagent. The coating reagent is an IgG polyclonal antibody, the washing solution is TBST, and its components For: Tris10mM, NaCl 150mM, Tween-20 0.05% (V / V), HCl to adjust the pH to 7.6, the capture reagent is biotin-coupled lamprey immune protein (LIP-Biotin), which can specifically recognize IgG in serum Glycoprotein Neu5Gc modification site, the detection reagent is streptavidin coupled with horseradish peroxidase (HRP), the chromogenic reagent is TMB chromogenic solution, the termination reagent is sulfuric acid solution, and biotin-streptavidin The Neu5Gc-modified IgG can be accurately quantified by the Hosin Reaction System.

[0053] Firstly, a standard curve for detecting Neu5Gc content is established, and the specific method is as follows:

[0054] (1) The pure product...

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Abstract

The invention discloses a liver cancer biomarker and a kit for detecting liver cancer, and belongs to the technical field of biomarkers and kits. Abnormal high expression of Neu5Gc modified IgG in serum of a liver cancer patient is found for the first time, the Neu5Gc modified IgG is used as a biomarker for diagnosing the liver cancer, the invention further provides a kit for detecting the liver cancer, the kit comprises a coating reagent, a cleaning solution, a capture reagent, a detection reagent, a color developing reagent and a termination reagent, the capture reagent is biotin-coupled lamprey immune protein, and the detection reagent is a biotin-coupled lamprey immune protein. The kit can specifically recognize the Neu5Gc modification site of IgG glycoprotein in serum, the detection reagent is streptavidin coupled with horse radish peroxidase (HRP), and the Neu5Gc modified IgG can be accurately quantified through a biotin-streptavidin reaction system. The kit disclosed by the invention can efficiently and specifically diagnose liver cancer, has monitoring efficiency on the effectiveness of treatment means of patients, and has indicating significance on prognosis monitoring of the patients.

Description

technical field [0001] The invention belongs to the technical field of biomarkers and kits, and in particular relates to a liver cancer biomarker and a kit for detecting liver cancer. Background technique [0002] Liver cancer is a cancer with a high mortality rate. Primary liver cancer or liver metastasis from other tumors can cause severe liver damage. Most primary liver cancers in adults belong to one of two types: hepatocellular carcinoma and cholangiocarcinoma / cholangiocarcinoma, which is one of the eight most common cancers in the world, and hepatocellular carcinoma is one of the common malignant tumors in my country, ranking first The death rate of malignant tumors ranks second in my country. The incidence of liver cancer in men is at least 2 to 3 times higher than that in women. The early symptoms are not obvious and atypical. The time between the beginning of tumor growth and the first symptoms is long, which is the main reason for the high mortality rate. It is in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57438G01N33/57488G01N33/68
Inventor 逄越李庆伟曹旭韩英伦
Owner LIAONING NORMAL UNIVERSITY
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