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Specific biomarker set for non-invasive diagnosis of liver cancer

A biomarker and specific technology, applied in disease diagnosis, biological testing, drug combination, etc.

Active Publication Date: 2016-02-10
DRAGON VICTORY DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Specific biomarker set for non-invasive diagnosis of liver cancer
  • Specific biomarker set for non-invasive diagnosis of liver cancer
  • Specific biomarker set for non-invasive diagnosis of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1a

[0064] Protein extraction from patient biopsies

[0065] Paired biopsies (tumor biopsies versus adjacent normal tissue) of 500 mg patients were collected and washed with PBS. Freeze the tissue by submersion in liquid nitrogen and immediately homogenize the tissue with a pestle and mortar. To the homogenized sample, a lysis solution (8M urea, 4% CHAPS, 2% IPG buffer, 0.2 mg / ml PMSF) was added, followed by vortexing for at least 5 min until the tissue was completely dispersed. The lysate was then clarified by centrifugation at 14,000 rpm for 10 minutes at 4°C. The supernatant was further washed by 2DCleanUpkit (Amersham) to remove salts and impurities. Resuspend the pellet with minimal volume of rehydration solution (without addition of DTT & IPG buffer). Protein concentrations were then measured by the Bio-Rad protein assay and aliquots of 200 g / tube were stored at -70°C.

Embodiment 1b

[0067] Protein Analysis by 2D Electrophoresis

[0068] To 1 ml rehydration stock solution was added 2.8 mg DTT, 5 μl pharmalyte or IPG buffer and 2 μl bromophenol blue. 50-100 μg protein sample was added to a 13 cm Immobiline Dry Strip (IPG strip) containing 250 μl rehydration solution. After removing the protective cover, position the IPG strips in the strip rack, gel side down, and cover with CoverFluid to prevent dehydration during electrophoresis. The strips were then placed on an Ettan IPGphor (Amersham) for isoelectric focusing (first dimension electrophoresis).

[0069] After the first dimension electrophoresis, the IPG strips were equilibrated with equilibration solution (6M urea 2% SDS, 50mM TrisHCl pH6.8, 30% glycerol, 0.002% bromophenol blue, 100mgDTT per 10ml buffer and 250mgIAA per 10ml buffer), and then used Wash 4–5 times with 1xSDS electrophoresis buffer. The IPG strip was placed on top of the second dimension gel and covered with sealing solution (0.5% low ...

Embodiment 2a

[0071] Example 2a (SEQ ID NO.1)

[0072] Amino acid sequence of Bmi1

[0073] MHRTTRIKITELNPHLMCVLCGGYFIDATTIIECLHSFCKTCIVRYLETSKYCPICDVQVHKTRPLLNIRSDKTLQDIVYKLVPGLFKNEMKRRRDFYAAHPSADAANGSNEDRGEVADEDKRIITDDEIISLSIEFFDQNRLDRKVNKDKEKSKEEVNDKRYLRCPAAMTVMHLRKFLRSKMDIPNTFQIDVMYEEEPLKDYYTLMDIAYIYTWRRNGPLPLKYRVRPTCKRMKISHQRDGLTNAGELESDSGSDKANSPAGGIPSTSSCLPSPSTPVQSPHPQFPHISSTMNGTSNSPSGNHQSSFANRPRKSSVNGSSATSSG

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Abstract

Cells within liver tumour mass comprise a unique set of proteins / tumour antigens when compared to the normal liver tissues epithelial cells juxtaposed to the tumour. The presence of tumour antigens couples the production of auto-antibodies against these tumour antigens. The present invention relates to the identification and elucidation of a protein set that can act as a novel marker set for liver cancer diagnosis and prognosis. Specifically, it relates to a kit that enables diagnostic and prognostic measurement of auto-antibodies in serum of liver cancer patients. The present invention provides a non-invasive, specific, sensitive, and cost effective detection and quantification method by evaluating a set of validated liver cancer proteins / tumour antigens, which includes Bmi-1, VCC1, SUMO-4, RhoA, TXN, ET-1, UBE2C, HDGF2, FGF21, LECT2, SOD1, STMN4, Midkine, IL-17A or IL26, to complement the conventional diagnostic methods.

Description

[0001] Copyright Notice / Permission [0002] A part of the disclosure content of this patent document contains content subject to copyright protection. The copyright owner has no objection to anyone's photocopying of the patent document or the patent disclosure as it appears in the Patent and Trademark Office patent file or records, but otherwise reserves all copyrights. The following statement applies to the procedures, experiments and data described below and in the figures accompanying this article: Copyright 2014, Vision Global Holdings Limited, all rights reserved. technical field [0003] The present invention describes a method for the detection and quantification of a specific and novel panel of hepatocellular carcinoma (HCC) tumor biomarkers by measuring the corresponding autoantibodies in the sera of liver cancer patients. The panel of biomarkers included Bmi1, VCC1, SUMO-4, RhoA, TXN, ET-1, UBE2C, HDGF2, FGF21, LECT2, SOD1, STMN4, midkine, IL-17A and IL26. More s...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68G01N33/58
CPCG01N33/57438G01N33/57488G01N33/582G01N33/6803G01N33/533G01N33/6863G01N33/6869G01N33/564G01N2800/56G01N2800/60A61P35/00G01N33/57484
Inventor 文咏贤卫凤文黄炳镠卫智贤
Owner DRAGON VICTORY DEV
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