Method for constructing hepatoma protein group fingerprint model and serum detection application thereof
A proteome and fingerprint technology, applied in the field of malignant tumor detection, can solve the problems of undetectable small liver cancer, and achieve the effects of easy detection, good repeatability, and prolonged survival
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Embodiment 1
[0029] Example 1 A serum proteome fingerprint model and construction method for detecting liver cancer according to the present invention:
[0030] 1. Serum Pretreatment
[0031] Serum samples were thawed in an ice bath and centrifuged at 3000r / min for 5 minutes, 6 μl of serum was added to 18 μl of urea, mixed thoroughly, ice bathed for 30 m, and diluted to 120 μl with WCX2 binding buffer.
[0032] 2. Pretreatment of WCX-2 protein chips
[0033] Pretreat the WCX-2 protein chip with 5 μl of 10 mmol / L HCI for 10 min, wash the chip with M-Q water for 3 times, install the chip on the Bioprocessor, add 150 uL of binding buffer (50 mmol / L sodium acetate, pH 4.0) to each point of the chip, Shake and incubate at room temperature for 5 min, discard the liquid, and repeat 1 time.
[0034] 3. Combination of chips and samples
[0035] Add 100 μl of the diluted sample to the chip and incubate with shaking. Binding buffer was added for elution twice in a row, 5 min each time. The Biopr...
Embodiment 2
[0042] Embodiment 2 Partial examples
[0043] 1. Reagents and chips
[0044] Acetonitrile, trifluoroacetic acid, urea, protease inhibitors, Hepes, Tris-HCl, CHAPS and OGP were purchased from Sigma. WCX2 protein chips were purchased from Ciphergen, USA. Protein standard Aβ-(1-42) was purchased from Sigma.
[0045] 2. Specimen source
[0046] Among the 160 serum samples, the serum samples of 60 patients with hepatocellular carcinoma were from newly diagnosed patients admitted to the surgical department of major hospitals in Guangzhou from August 2002 to August 2003; Volunteer healthy people, observed for half a year and no tumor occurred. 60 cases of liver cancer were diagnosed as hepatocellular carcinoma by postoperative pathology. There were 85 males and 15 females with hepatocellular carcinoma, aged from 32 to 75 years old, with an average of 59.6 years old. According to the "Clinical Diagnosis and Staging Standards for Primary Liver Cancer" of the Liver Cancer Professi...
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