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A method for detecting tumor-related markers in serum of patients with esophageal cancer

A tumor-related, patient serum technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as inability to produce antibodies

Inactive Publication Date: 2011-11-30
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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Problems solved by technology

Since SEREX needs to establish an expression library, its analysis also has limitations in the genetic material of the tumor individual genome.
In addition, SEREX technology cannot generate antibodies against protein isoforms or different post-translational modified forms of proteins

Method used

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  • A method for detecting tumor-related markers in serum of patients with esophageal cancer
  • A method for detecting tumor-related markers in serum of patients with esophageal cancer
  • A method for detecting tumor-related markers in serum of patients with esophageal cancer

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Embodiment 1

[0019] see figure 1 with figure 2 A method for detecting tumor-associated markers in the serum of patients with esophageal cancer, the steps of which are as follows:

[0020] A. Collect peripheral blood samples from patients with esophageal cancer, centrifuge at 4°C within 2-3 hours after isolation, and separate the upper serum to obtain tumor serum;

[0021] B. Collect peripheral blood samples from healthy people, centrifuge at 4°C within 2-3 hours after isolation, and separate the upper serum to obtain control serum;

[0022] C, extract the total protein of cell lines cultured in vitro; the specific method of the present embodiment is: for the esophageal cancer cell line EC-0156 cultured in vitro, the culture solution in the culture dish is discarded, and pre-cooled 1 × phosphate buffered saline ( PBS), pH 7.4 (137mM NaCl, 2.7mM KCl, 10mM NaCl 2 HPO 4 , 2mM KH 2 PO 4 ), washed 3 times, added appropriate amount of protein lysate (50mM Tris-HCl, pH7.4, 150mM NaCl, 1% Tr...

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Abstract

The invention relates to a method for detecting relevant tumor markers in the serum of patients with esophageal cancer. Serum is used as a molecular probe to immunoprecipitate potential tumor antigens in tumor cells, and the enriched tumor antigen-antibody complex is subjected to SDS-PAGE Electrophoretic separation, Coomassie brilliant blue staining, enzymatic digestion of specific protein bands, peptide extraction, reversed-phase high-performance liquid chromatography (RP-HPLC) separation and electrospray ionization coupled with online ion trap mass spectrometry (ESI-IT- MS) identification and other comprehensive technical routes to screen serum marker proteins for esophageal cancer. The screened Ku70 protein was confirmed as a new candidate marker for esophageal cancer by immunoblotting, and the significantly increased expression of Ku70 protein was related to the carcinogenesis of early esophageal cancer.

Description

technical field [0001] The invention relates to a method for detecting tumor-related markers in serum of patients with esophageal cancer. The method uses the serum of patients with esophageal cancer as antibodies to capture tumor antigens, and then enriches tumor antigen-antibody complexes. The enriched tumor antigen-antibody complexes are separated by SDS-PAGE electrophoresis and identified by mass spectrometry to obtain tumor markers. Background technique [0002] Esophageal cancer is one of the common malignant tumors in my country, and there is still a lack of effective early diagnosis methods. In the process of tumor development, the instability of the genome will cause many protein expression and degradation disorders. These abnormally changed proteins or small molecular peptides can be recognized by the body's immune system as "non-self" substances (self-antigens) and induce the body to produce anti-antigens. Humoral immune response to tumor autoantigens, with autoan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88G01N33/574G01N33/561G01N33/531
Inventor 赵晓航何洪智吴旻
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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