Protein complex and application of protein complex in tumor diagnosis and/or prognosis evaluation
A technology for protein complexes and prognosis assessment, applied in hybrid peptides, biomaterial analysis, chemical instruments and methods, etc., can solve the problem of not finding cancer markers
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Embodiment 1
[0083] Embodiment 1 (preparation embodiment)
[0084] Separation of protein complexes in serum, separation and identification of protein complex components
[0085] The study subjects were 1591 serum samples, including 501 healthy controls, 282 lung cancer, 209 colorectal cancer, 201 pancreatic cancer, 205 gastric cancer and 193 thyroid cancer patients.
[0086] 1. Native-PAGE separation method of protein complex in serum
[0087] The composition and electrophoresis conditions of Native-PAGE gel are as follows:
[0088] Gradient separation gel (4%-10% (mass volume ratio), acrylamide / methylenebisacrylamide=37.5:1 (mass ratio)) components are prepared from heavy liquid and light liquid through gradient mixer. Heavy liquid (10% (mass volume ratio), acrylamide / methylenebisacrylamide=37.5:1 (mass ratio)) composition is: TBM (90mM tris hydroxymethyl aminomethane (Tris base) / 90mM boric acid (Boricacid) / 1.5mM magnesium chloride (MgCl 2 )), 0.05% (volume ratio) tetramethylethylene...
Embodiment 2( application Embodiment 1
[0126] For the application of Native-PAGE to separate protein complexes in serum, the specific information of the serum samples used is as follows (Table 2).
[0127]
[0128] 1. Selection and evaluation of internal reference
[0129] (1) Serum samples from 1591 cases were collected, including 282 cases of lung cancer (lung cancer, LC), 209 cases of colorectal cancer (colorectal cancer, CC), 201 cases of pancreatic cancer (pancreatic cancer, PC), and 205 cases of gastric cancer (stomach cancer, SC), 193 cases of thyroid cancer (thyroid cancer, TC) and 501 cases of healthy controls (control), carry out Native-PAGE electrophoresis according to the method in Example 1, a part of mixed healthy human serum is used as external reference (N), The purpose is to reduce the impact on the optical density value of the gel band during the process of electrophoresis, staining and gel scanning, and provide accurate data for subsequent data analysis. Take 2 μL serum sample and 2 μL extern...
Embodiment 3( application Embodiment 2
[0163] Use SPSS software to draw a single protein complex and comprehensively use the receiver operating curve (ROC) of the relative content of each protein complex
[0164] According to the equation in embodiment 2 step 2 (6), draw the ROC curve, calculate the area under the ROC curve (AUC) and 95% credible interval (95%CI), the threshold (cut-off value) of inspection, sensitivity ( sensitivity) and specificity (specificity), etc. The analysis results showed that the sensitivity and specificity of lung cancer, colorectal cancer, pancreatic cancer, gastric cancer, and thyroid cancer were all greater than 80.00% compared with the healthy control group. The sensitivity and specificity of the diagnosis when the relative content of each related target protein complex is detected jointly is higher than that of any single target protein complex. The present invention only lists the results after joint detection of each relevant index (see Table 6).
[0165] Table 6 ROC curve analy...
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