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Detection method of IgG (immunoglobulin G) core fucosylation level and use thereof

A fucosylation and horizontal technology, applied in the field of protein, can solve the problems of clinical application limitations, increasing the difficulty of diagnostic marker research, difficult dynamic observation, etc.

Active Publication Date: 2011-09-07
HEPATOBILIARY SURGERY HOSPITAL SECOND MILITARY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Liver cirrhosis is the main adverse outcome of a variety of chronic liver diseases, especially viral hepatitis; the gold standard for its diagnosis is still liver biopsy pathological diagnosis, but it is well known that liver biopsy is an invasive examination, difficult to observe dynamically, and its clinical application is limited
The formation of liver cirrhosis is a complex process involving multiple factors and multiple cells, which further increases the difficulty of the study of its diagnostic markers
[0005] Clinical studies have confirmed that patients with liver disease often have elevated levels of immunoglobulin, but their structural characteristics and their significance in the development of liver disease have not been reported

Method used

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  • Detection method of IgG (immunoglobulin G) core fucosylation level and use thereof
  • Detection method of IgG (immunoglobulin G) core fucosylation level and use thereof
  • Detection method of IgG (immunoglobulin G) core fucosylation level and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Detection and analysis of IgG core fucosylation levels in normal control group (Control), chronic hepatitis B group (CHB) and HBV cirrhosis (LC) group.

[0028] Serum samples from 20 cases of CHB and LC patients who came to our hospital for treatment from September 2006 to September 2009 were collected, including 16 males and 4 females in each group; the average age was 51.6±13.2 years old. All selected patients must also meet the following criteria: (1) Hepatitis B patients who are infected with hepatitis B virus (HBV); TP, ALB and IgG do not have the following rules, so exclude human immunodeficiency virus, hepatitis C, D, E and syphilis and other pathogens other than HBV infection; (3) exclude autoimmune liver disease, alcohol liver disease, drug-induced liver disease, and Wilson disease. In addition, 20 healthy subjects were selected as the control group, with an average age of 51.2±10.3 years. There was no statistically significant difference in gender...

Embodiment 2

[0046] Embodiment 2: Detect patient serum TP, IgG, LCA-TP and LCA-IgG

[0047] Specific experimental steps:

[0048] 1) Blood was drawn from patients 1, 2, 3 and 4, and serum was collected.

[0049] 2) Separation and purification of core fucosylated total protein

[0050] a. Add 200ul serum to 300ul cleaning solution, mix well and set aside;

[0051] b. Put the prepacked LCA lectin affinity column into the outer tube of the spin column, room temperature 3000rpm / min, 20 seconds;

[0052] c. Add the diluted serum to the prepacked LCA lectin column, let it stand for 10 minutes, 3000rpm / min, 20 seconds;

[0053] d. Add 1ml cleaning solution, 3000rpm / min, 20 seconds;

[0054] e. Put the prepacked LCA lectin column into the outer tube of the new spin column, add 500ul eluent, room temperature 3000rpm / min, 20 seconds;

[0055] f. Collect the liquid in the tube.

[0056] 3) Detection and analysis

[0057] A Hitachi 7170 biochemical analyzer was used to detect TP, ALB and IgG in...

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Abstract

The invention belongs to the technical field of proteins, and relates to the detection of glycoprotein, specifically to a detection method of the IgG core fucosylation level and a use thereof, wherein the method comprises the step of calculating the IgG amount of the core fucosylation / the glycoprotein total amount of the core fucosylation. The method can not distinguish healthy persons from hepatitis B patients, but can distinguish whether the diagnosed hepatitis B patient is a chronic hepatitis patient or a cirrhosis patient.

Description

technical field [0001] The invention belongs to the technical field of proteins and relates to the detection of glycoproteins, in particular to a method for detecting the fucosylation level of the core of immunoglobulin IgG and its application. This method, although unable to distinguish between healthy and hepatitis B virus (HBV)-infected patients, can be used to distinguish chronic hepatitis patients from those with cirrhosis among those diagnosed with HBV infection. Background technique [0002] In recent years, people have gradually realized that protein glycosylation modification plays an important role in the occurrence and development of many diseases. The liver is an important site for protein synthesis and processing, which is closely related to the glycosylation state of serum proteins. In our previous studies, we have found characteristic N-glycan structural markers in the blood of patients with HBV infection, liver cirrhosis and liver cancer. However, there is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48
Inventor 高春芳衣常红房萌赵云鹏顾星季君王坤
Owner HEPATOBILIARY SURGERY HOSPITAL SECOND MILITARY MEDICAL UNIV
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