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Sugar chain marker for diagnosing PBC patients with positive and negative anti-ACA antibodies and application of sugar chain marker

A technology of markers and antibodies, applied in disease diagnosis, biological testing, biomaterial analysis, etc., can solve the problem of loss of IgG pro-inflammatory activity and anti-inflammatory activity

Pending Publication Date: 2021-08-20
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, complete removal of polysaccharides leads to loss of IgG pro-inflammatory and anti-inflammatory activities

Method used

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  • Sugar chain marker for diagnosing PBC patients with positive and negative anti-ACA antibodies and application of sugar chain marker
  • Sugar chain marker for diagnosing PBC patients with positive and negative anti-ACA antibodies and application of sugar chain marker
  • Sugar chain marker for diagnosing PBC patients with positive and negative anti-ACA antibodies and application of sugar chain marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Lectin microarray analysis of serum IgG glycosylation

[0034] Experimental specimens: PBC patients in this study: anti-ACA antibody positive (20 cases), anti-ACA antibody negative (20 cases), see Table 1. The diagnosis of PBC patients were in line with the diagnostic criteria of the corresponding disease. Fresh blood was collected from all the people enrolled in the group, and the serum was separated immediately and frozen at -80°C for later use.

[0035] Table 1. Basic information of the research objects in the lectin chip experiment

[0036]

[0037] A lectin microarray containing 56 lectins was used to detect the glycosylation status of experimental samples. Lectins can specifically bind to glycan molecules at the end of glycoproteins to form complexes. Through the specific binding of different lectins to glycans, the type and content of glycans on the surface of the target protein can be studied. Lectin microarray has been more and more widely used ...

Embodiment 2

[0046] Example 2 Serum lectin imprint verification experiment

[0047] Experimental specimens and methods: In order to further clarify the reliability of the above-mentioned lectin microarray detection conclusions, the same batch of anti-ACA antibody positive and negative patients (6 cases each) and the new batch of anti-ACA antibody positive and negative patients were tested by lectin microblotting. Negative patients (10 cases each) were verified by lectin blot, see Table 4. Serum samples were diluted 1:100, mixed with sample buffer, boiled at 100°C for 5 minutes, performed SDS-PAGE electrophoresis in 10% precast gel, and then electrotransferred the protein in the precast gel to PVDF membrane. After the successfully transferred PVDF membrane was blocked, it was hybridized with cy3-labeled lectin, and finally the fluorescence signal was detected by a fluorescence imager. The intensity of the fluorescent signal is directly proportional to the binding force of the glycoprotein ...

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Abstract

The invention discloses a sugar chain marker for diagnosing anti-ACA antibody positive and negative PBC patients and application of the sugar chain marker. A lectin chip containing 56 types of lectin is adopted to detect the glycan difference of specific binding between serum IgG of an anti-ACA antibody positive patient and an anti-ACA antibody negative patient in the PBC and the lectin. The results show that the content of PSA and Con A lectin binding glycans is reduced in the anti-ACA antibody positive patients compared to that in the negative patients. Since PSA and Con A lectin are specific binding mannose, this indicates that the expression of mannose levels in the anti-ACA antibody positive patients is reduced. A lectin immunoblotting verification result also shows that the content of the PSA and Con A lectin binding glycans is still reduced in the anti-ACA antibody positive patients. Therefore, the compound formed by combining the PSA lectin and the IgG and / or the compound formed by combining the Con A lectin and the IgG can be used as a carbohydrate chain marker for diagnosing the anti-ACA antibody positive PBC patients and the anti-ACA antibody negative PBC patients.

Description

technical field [0001] The invention belongs to the field of biological detection, and specifically relates to a sugar chain marker for diagnosing anti-ACA antibody-positive PBC patients and anti-ACA antibody-negative PBC patients and its application. Background technique [0002] Primary biliary cholangitis (PBC) is a chronic autoimmune liver disease that can develop into cirrhosis and eventually liver failure. Disease development is highly variable and can extend over decades. In addition to the serological marker AMA of PBC, the positive rate of ANA (antinuclear antibody) in PBC patients can reach 50%. ANA generally refers to a class of autoantibodies against eukaryotic nuclear components, which can appear in a variety of autoimmune diseases. In addition to PBC-specific autoantibodies ANA (SP100 and gp210), approximately 25% of PBC patients are positive for anticentromere antibodies (ACA). The target antigen of the anti-ACA antibody is a 17-19kD protein in the centrome...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68G01N2800/08
Inventor 胡朝军曾小莉唐诗逸李晞李斯亭李梦涛曾小峰
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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