Screening method, application and kit for disease-related markers

A technology of markers and complexes, applied in the fields of molecular biology and immunology, can solve the problem of lack of serological markers and achieve the effect of narrowing the scope of analysis

Active Publication Date: 2022-05-20
PEKING UNIV
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the diagnosis of severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) virus infection mainly relies on nucleic acid detection, there is still a lack of serological markers, and there is no feasible method for the degree of tissue damage. Sure

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Screening method, application and kit for disease-related markers
  • Screening method, application and kit for disease-related markers
  • Screening method, application and kit for disease-related markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening of potential markers related to new coronary pneumonia

[0052] The first part, purification of immunoglobulin complexes and protein profile analysis.

[0053] Take 200 µL of serum from 9 patients with new coronary pneumonia (2 weeks of recovery) and 9 healthy people, dilute it 10 times with PBS, and then use Jacalin, Protein G, and anti-human IgM to affinity-purify the serum through an affinity chromatography column The IgA complex, IgG complex and IgM complex, these three Ig complexes are then used for protein spectrum sequencing analysis, the operation process is as follows figure 1 shown.

[0054] Jacalin can affinity-purify IgA in serum, and then purify various proteins that bind to IgA together. Protein G can affinity-purify IgG in serum, and then purify various proteins that bind to IgG together. Anti-human IgM can affinity-purify IgM in serum, and then purify various proteins that bind to IgM together.

[0055] After further analysis of t...

Embodiment 2

[0078] Example 2 Screening of potential markers related to coronary heart disease (CHD) and acute myocardial infarction (AMI)

[0079] Part I: Screening of potential markers related to coronary heart disease (CHD)

[0080] Purification of IgG complexes and protein profile analysis: take the serum of patients with coronary heart disease (CHD) and normal people (NM), dilute it with PBS, and then use Protein G to affinity-purify the serum in the serum through an affinity chromatography column. IgG complex. Mass spectrometric analysis of proteins bound to IgG.

[0081] For protein profiling results see Figure 4 , the protein names represented by different protein markers in the figure are shown in the table below:

[0082] Numbering Gene English full name Chinese full name P02679 FGG Fibrinogen gamma chain fibrinogen gamma chain P02671 FGA Fibrinogen alpha chain fibrinogen alpha chain P08493 MGP Matrix Gla protein extracellular mat...

Embodiment 3

[0096] Example 3 Screening of glioma-related markers

[0097] Purification of Ig complexes and protein profile analysis: Sera from patients with glioma, coronary heart disease, lung cancer, and gastrointestinal cancer were in four groups, with 10 people in each group. The serum was diluted with PBS and passed through Protein G. Affinity chromatography column for affinity purification of IgG complexes in serum. The proteins in the resulting IgG complexes were subjected to mass spectrometry as well as glycan profiling.

[0098] see Figure 8 and Figure 9 , the results of mass spectrometry showed that, compared with other groups, two proteins, Protein S100-A8 (5 / 10) and Fibronectin (fibronectin) (3 / 10), were higher in IgG complexes purified from blood samples from patients with glioma. horizontal expression. see Figure 10 and Figure 11 , Suprabasin (5 / 10, that is, 5 out of 10 samples showed high expression) and Annexin A2 (annexin A2) (4 / 10) appeared at high frequency in...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a screening method, application and kit of disease-related markers. The screening method includes: using substances capable of binding immunoglobulin (Ig), purifying Ig complexes from the first batch of healthy sample sera and diseased sample sera; performing protein spectrum sequencing analysis on proteins that bind to Ig, and comparing The difference between healthy samples and diseased samples, finding the differential protein that only appears in diseased samples, is the potential marker related to the disease. In addition, the potential marker can be further verified by the following steps: use the second batch of serum from healthy samples and diseased samples (extended case), purify the Ig complex, and use the specific antibody of the differential protein for further identification. This method first obtains Ig complexes from serum (instead of whole serum), and then performs protein spectrum sequencing combined with specific antibody analysis on the proteins that bind to Ig, which can quickly and effectively screen out disease-related markers.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and immunology, in particular to a screening method, application and kit for disease-related markers. Background technique [0002] A biomarker refers to a substance in a living body that can be objectively detected and evaluated, and can be used as an indicator of normal biological processes, pathological processes, or pharmacological responses to therapeutic interventions. In the research of disease diagnosis and treatment, finding and discovering valuable biomarkers has become a research hotspot in the current medical field. [0003] The patent with publication number CN108956791A discloses a method for large-scale screening of protein biomarkers: homogenize and lyse biological tissue control samples and reference samples (pathological samples); MS / MS liquid chromatography-mass spectrometry is used for protein identification; by comparing the number of mass spectrometry analysis tim...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/573G01N33/92
CPCG01N33/6854G01N33/686G01N33/6893G01N33/57423G01N33/57407G01N33/57488G01N33/573G01N33/86G01N33/92G01N2800/12G01N2800/26G01N2800/324
Inventor 邱晓彦段富刚朱珠戴慧张奕潇王一凡姜文华
Owner PEKING UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap