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The use of granulin-epithelin precursor (GEP) antibodies for detection and suppression of hepatocellular carcinoma (HCC)

A hepatocellular carcinoma and antibody technology, applied in the direction of antibodies, immunoglobulins, animal/human proteins, etc., can solve the problems of limited targeted therapy, unsuitable treatment form, poor prognosis, etc.

Active Publication Date: 2009-10-07
THE UNIVERSITY OF HONG KONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prognosis of HCC patients with hepatocellular carcinoma is generally poor, with a median survival time of less than 1 year, because most hepatocellular carcinomas are unresectable, not suitable for new treatment modalities, and the efficacy of chemotherapy is low
However, the development of targeted therapies for HCC, including antibody therapy, is limited, thus, new therapeutic targets are urgently needed
[0009] The diagnostic significance of serum GEP in any human cancer has not been reported to date

Method used

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  • The use of granulin-epithelin precursor (GEP) antibodies for detection and suppression of hepatocellular carcinoma (HCC)
  • The use of granulin-epithelin precursor (GEP) antibodies for detection and suppression of hepatocellular carcinoma (HCC)
  • The use of granulin-epithelin precursor (GEP) antibodies for detection and suppression of hepatocellular carcinoma (HCC)

Examples

Experimental program
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Effect test

Embodiment 1

[0048] patient sample

[0049] The study protocol was approved by the Institutional Review Board of The University of Hong Kong, and signed consent forms were obtained from patients and control subjects. Between March 1999 and October 2004, 107 patients diagnosed with primary HCC, 38 patients with chronic hepatitis B (only those with no signs of malignant disease during the follow-up period of more than 2 years) were included in this study) and 72 healthy donors negative for hepatitis B surface antigen (HBsAg) were obtained. Serum HBsAg was positive in 96 (89.7%) HCC patients, so the control group included chronic hepatitis B patients and healthy volunteers. Serum samples were frozen at -70°C until use. Tumor and adjacent non-tumor liver tissue were collected from HCC patients, snap frozen in liquid nitrogen, and stored at -70°C until use. Parallel sections were formalin-fixed and embedded in paraffin for histological examination and immunohistochemical studies. Clinical a...

Embodiment 2

[0051] cell line

[0052] Human HCC cell lines Hep3B, HepG2 and Huh7 (American Tissue Culture Collection (American Tissue Culture Collection), Manassas, VA) and Japan Health Science Research Resources Bank ((Japan Health Science Research Resources Bank), Osaka, Japan) ) were maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (Gibco BRL, Carlsbad, CA).

Embodiment 3

[0054] Build up antibodies

[0055] by conjugating the keyhole with 33 μg A custom GEP-specific peptide of hemocyanin (KLH), SEQ ID No: 3, was immunized subcutaneously in BALB / c mice with complete Freund's adjuvant (Sigma-Aldrich, Dorset, UK) to generate GEP-specific antibodies. For subsequent booster immunizations, the same amount of antigen in Freund's incomplete adjuvant was injected intraperitoneally twice a week. After each booster immunization, serum antibody activity against the immunizing antigen was monitored using an ELISA against the peptide antigen. Mice showing high serum antibody titers against the antigen were given a booster immunization with the last intravenous injection of the antigen, and spleens were harvested 3 days later.

[0056] Production of anti-GEP monoclonal antibody A23

[0057] Spleens were collected from mice showing high antibody titers in their sera. Fusion of splenocytes with the non-productive myeloma cell line NS0 was performed followi...

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Abstract

This invention provides methods for detecting serum GEP level. This invention further provides methods for determining whether a subject is afflicted with Hepatocellular carcinoma (HCC) by measuring serum GEP level. In another embodiment, this invention provides methods for the suppression of HCC growth and progression both in vitro and in vivo by treating a patient with anti-GEP monoclonal antibody A23.

Description

[0001] Cross References to Related Applications [0002] This application is a continuation-in-part of US Patent Application Serial No. 10 / 836,390 filed April 29,2004. This application also claims priority to US Provisional Patent Application No. 60 / 861,318, filed November 28, 2006. The entire content of each of the foregoing applications is incorporated by reference into this application. field of invention [0003] The present invention relates to granulin-proepithelin (GEP) and methods of affecting the expression, translation and biological activity of GEP in hepatocellular carcinoma (HCC). Another aspect of the present invention relates to the detection method of GEP, which is a possible method for the diagnosis and treatment of HCC. [0004] Several publications are referred to in this article with Arabic numerals in parentheses. Full citations for these references can be found at the end of the specification preceding the claims. The entire contents of these publica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/535G01N33/577C07K14/00A61K39/395A61P35/00
CPCC07K16/22C07K14/475A61K2039/505G01N33/57438C07K2317/73A61P35/00
Inventor 张兆恬何颂怡范上达
Owner THE UNIVERSITY OF HONG KONG
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