Group of specific polypeptides and application thereof in preparing reagents for early diagnosis of lung cancer
A specific and versatile technology, applied in the field of preparation of early diagnostic reagents for lung cancer, to achieve huge market prospects and social and economic benefits
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Embodiment 1
[0014] Example 1 Extraction of Serum Polypeptides from Metallic Copper Chelated Nano Magnetic Beads
[0015] 1. Materials:
[0016] Binding buffer solution: 0.1mol / l phosphate buffer solution, pH 4
[0017] Wash buffer solution: 0.1% formic acid in water
[0018] Eluent: 0.1% formic acid and 50% acetonitrile in water
[0019] Serum of 254 cases of lung cancer patients
[0020] 257 cases of control serum
[0021] Metal copper chelated nano magnetic beads: (self-made, patent application number: 200610002030.1)
[0022] 2. Method results:
[0023] (1) Preparation of metal copper chelated nano magnetic beads:
[0024] 1. Materials:
[0025] FeCl3: Analytical pure AR, batch number QD, March 11, 2004, produced by Sinopharm Chemical Reagent Co., Ltd.; PEG 1500: Mr1400-1600, batch number: 25300-68-3, FLUKA product; pyrrolidone (2-pyrrolidone): for synthesis, S34844146, product of MERCK-Schuchardt; diethyl ether: analytically pure, batch number: 20050315, produced by Tianjin Ch...
Embodiment 2
[0036] Example 2 Bioinformatics analysis of serum polypeptide map of lung cancer
[0037] 1. Software:
[0038] Biosun analysis software SVM package, http: / / www.csie.ntu.edu.tw / ~cjlin / libsvm
[0039] 2. Method
[0040] Sera from 254 cases of lung cancer and 257 cases of controls (healthy volunteers, pneumonia, chronic obstructive emphysema, COPD, benign lung tumors) were randomly selected to establish a lung cancer diagnostic model and screen for differential peptides. The cross validation and recognition capability of the obtained diagnostic model are both above 95%, and the remaining tumors and control cases are used for verification, and the diagnostic accuracy is above 90%.
Embodiment 3
[0041] Example 3 Sequence Identification of Lung Cancer Specific Serum Polypeptides
[0042] 1. Materials:
[0043] Serum from 10 cases of lung cancer patients; 10 cases of serum from healthy controls;
[0044] Magnetic beads (self-made, patent application number: 200610002030.1); 9.4 TQ-FT-ICR-MS (Bruker Company).
[0045] 2. Method results:
[0046] 1. Referring to Example 1, the serum peptides were extracted with magnetic beads, and the peptides extracted from 10 cases of lung cancer were mixed, and the peptides extracted from 10 cases of normal controls were mixed. The mixed peptide samples were concentrated in a centrifugal dryer to remove the organic phase. Prepare for mass spectrometry analysis to identify marker peptide sequences.
[0047] 2. The peptide sample was injected into nano LC-9.4T Q-FT-ICR-MS, gradient eluted, and the peptide was identified by tandem mass spectrometry, and the obtained spectrum was analyzed by DataAnalysis software (Bruker Company) to ob...
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