Fingerprint model for liver cancer serum characteristic protein detection and preparation method thereof

A fingerprint and characteristic protein technology, which is applied in the field of protein fingerprint detection, can solve the problems that have not been reported, achieve the effect of comprehensive population range, overcome poor repeatability and low stability, and reduce the fatality rate

Inactive Publication Date: 2013-11-06
FUZHOU GENERAL HOSPITAL OF NANJING MILITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, based on the principle of glycoproteomics, the application of CLINPROT technology to establish the fingerprint model of liver cancer serum fucosylation abnormal glycoproteome has not been reported at home and abroad so far.

Method used

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  • Fingerprint model for liver cancer serum characteristic protein detection and preparation method thereof
  • Fingerprint model for liver cancer serum characteristic protein detection and preparation method thereof
  • Fingerprint model for liver cancer serum characteristic protein detection and preparation method thereof

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preparation example Construction

[0023] The present invention is used for the preparation method of the fingerprint atlas model that detects liver cancer serum characteristic protein, comprises the following steps:

[0024] 1) Serum samples from patients with primary liver cancer related to hepatitis B virus that were clearly diagnosed by pathological and DNA examinations were collected as serum samples of the liver cancer group; serum samples from patients with liver cirrhosis caused by confirmed hepatitis B virus were confirmed Serum from patients with chronic hepatitis B without liver cirrhosis and those from healthy subjects determined by physical examination were used as serum specimens of the non-liver cancer group, and were cryogenically frozen for future use.

[0025] 2) MB-LAC LCA magnetic beads, which can specifically adsorb fucosylated abnormal glycoproteins, were used to adsorb the glycoproteins of the serum samples of the above-mentioned liver cancer group and non-liver cancer group, and then the ...

Embodiment 1

[0035] Take 2ml of venous blood from the patient to be tested without anticoagulation. Place at room temperature for 30 min to 1 h. Centrifuge at room temperature at 2500 rpm for 5 min, take equal aliquots of serum and store in a -80°C refrigerator. During the experiment, the specimens were taken out from the -80°C refrigerator and thawed in ice for later use. Take out one tube of MB-LAC LCA magnetic bead suspension from the 4°C refrigerator and pipette carefully to suspend the magnetic beads completely and evenly in the liquid phase; place the 200 μl sample tube on the well plate, add 2 μl magnetic beads ( MB) and 100 μl magnetic bead washing buffer (WB1), transfer the sample tube to the magnetic rack, let the magnetic beads adhere to the wall for 20 s, separate the magnetic beads from the liquid, and suck up the liquid after the liquid is clarified; Aspirate the supernatant; repeat once to ensure that the suspension is completely absorbed; put the sample tube on the orifice...

Embodiment 2

[0037] Take 2ml of venous blood from the patient to be tested without anticoagulation. Place at room temperature for 30 min to 1 h. Centrifuge at room temperature at 2500 rpm for 5 min, take equal aliquots of serum and store in a -80°C refrigerator. During the experiment, the specimens were taken out from the -80°C refrigerator and thawed in ice for later use. Take out one tube of MB-LAC LCA magnetic bead suspension from the 4°C refrigerator and pipette carefully to suspend the magnetic beads completely and evenly in the liquid phase; place the 200 μl sample tube on the well plate, add 2 μl magnetic beads ( MB) and 100 μl magnetic bead washing buffer (WB1), transfer the sample tube to the magnetic rack, let the magnetic beads adhere to the wall for 20 s, separate the magnetic beads from the liquid, and suck up the liquid after the liquid is clarified; Aspirate the supernatant; repeat once to ensure that the suspension is completely absorbed; put the sample tube on the orifice...

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Abstract

The invention relates to the field of protein fingerprint detection technology, especially to a fingerprint model for liver cancer serum characteristic protein detection. Seven abnormal fucosylation glycoproteins which include four up-regulated proteins and three down-regulated proteins are screened from serum as a characteristic proteome. On the base of the characteristic proteome, the fingerprint model for liver cancer serum characteristic protein detection is built. The molecular weights of the four up-regulated proteins are 9290.06 Dalton, 2660.24 Dalton, 2764.18 Dalton and 1862.71 Dalton respectively. The molecular weights of the three down-regulated proteins are 4282.99 Dalton, 3963.52 Dalton and 6023.31 Dalton respectively. The fingerprint model provides a new method for early diagnosis of liver cancer, provides a foundation for further finds of new tumor markers, and is beneficial to improve the accuracy of diagnosis of liver cancer.

Description

technical field [0001] The invention relates to the technical field of protein fingerprint detection, in particular to a fingerprint model for detecting liver cancer serum characteristic protein and a preparation method thereof. Background technique [0002] my country is one of the countries with high hepatitis B virus (HBV) infection. The mortality rate of liver cancer caused by HBV is high, and the 5-year survival rate after surgical treatment is low. An effective means to significantly improve the therapeutic effect is early diagnosis. At present, the commonly used method for primary detection of liver cancer is the determination of serum AFP content, but its sensitivity and specificity are not high. The study found that the serum AFP protein of liver cancer was highly fucosylated with high specificity. Detection of this part of AFP (AFP-L3) can greatly improve the diagnosis of liver cancer. However, in view of the negative serum AFP value of about half of patients w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62
Inventor 廖剑殷正丰兰小鹏钱海华王开宇
Owner FUZHOU GENERAL HOSPITAL OF NANJING MILITARY COMMAND P L A
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