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
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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...
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