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Biomarkers for liver diseases and method for using the same

a liver disease and liver disease technology, applied in the field of liver diseases, can solve the problems of reducing the activity of immune cells, prone to developing immune diseases, and people with impaired immune functions to develop immune diseases, and achieve the effect of greater accuracy and sensitivity

Inactive Publication Date: 2005-06-23
IND TECH RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0017] This invention is based on the use of autoantigen screening method, comprising the steps of: firstly purifying antibodies from normal persons, liver cirrhosis patients, and liver cancer patients respectively and immobilizing them in different columns; passing the cell extracts from liver disease related cell lines (HepG2 C3A & SNU-387) in sequence through the normal antibody column and patient antibody column to obtain autoantigens associated with liver cirrhosis and liver cancer; using those autoantigens as biomarker kits coupled with enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), or immunofluorescence to detect the presence of autoantibodies against said autoantigens in the screened specimen, and based on which, to determine whether the patient has liver cirrhosis or liver cancer. Since those biomarkers are identified based on existing autoantibodies, they can be developed into diagnostic kits to determine if the patient has such diseases based on the presence of autoantibodies against the biomarkers. Such method is much easier than direct screening of the antigen and offers greater accuracy and sensitivity.

Problems solved by technology

People with impaired immune functions are prone to develop immune diseases.
The first is reduced immunity, lower activity of immune cells, or reduced quantity of immune cells, such that the human body cannot fight off the invading bacteria, virus or mold, and becomes susceptible to contagious diseases, such as common cold, flu, pneumonia, enteritis, or even hepatitis and AIDS.
When real pathogens such as bacteria, virus or mold attack the human body at this time, the immune system is no longer able to put up resistance.
Such immune diseases arise from our own immune system having an identification problem that autoantibodies are produced against human body's own cells, resulting in tissue damage and illnesses.
But the biomarkers disclosed in those patents lack accuracy or are susceptible to interference to a certain extent.

Method used

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  • Biomarkers for liver diseases and method for using the same
  • Biomarkers for liver diseases and method for using the same

Examples

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example 1

Screening of Autoantigens Using Autoantibodies in Sera of Patients with Liver Diseases Purification of Autoantibodies in the Serum Sample

[0031] Firstly obtaining a serum of a patient with liver cirrhosis or liver cancer, diluting the serum with a binding buffer (20 mM PBS, pH 7.0) at the ratio of 1:10, and then filtering the diluted serum using a 0.45 μm filter membrane to prevent the blockage of column in subsequent steps; next rinsing a Protein G affinity column with the binding buffer ten times the column volume at the rate of 1 ml / min, and then passing the filtered serum sample over the Protein G affinity column at the rate of 0.2 ml / min to retain the antibodies in the column through affinity; rinsing the Protein G affinity column again using the binding buffer 5-10 times the column volume at the rate of 1 ml / min to remove substances in the serum sample that do not form affinity bonding with the column. Eluting antibodies from the column using an elution buffer (0.1 M Glycine-...

example 2

Determining the Availability of Autoantigens Identified by the Autoantigen Screening Method

[0063] To demonstrate the availability of 24 autoantigens identified in Example 1, further assay of serum samples from normal persons, liver cirrhosis patients and liver cancer patients using immunoassay (ELISA, RIA or immunofluorescence) and the aforesaid 24 biomarkers is carried out. The assay method includes the following steps as shown in FIG. 2: providing a specimen; using the biomarker selected from any one of the amino acid sequences with SEQ ID NO:1 to SEQ ID NO:24 or derivatives or fragments or variants or the combination thereof to capture the autoantibody in the specimen; and detecting the auto antibody.

[0064] In the example of enzyme-linked immunosorbent assay (ELISA), the following steps are taken: firstly diluting the biomarker with a coating buffer (choice of a. 50 mM Na2HCO3, pH=9.6, or b. 20 mM Tris-HCl, pH=8.5, or c. 10 mM PBS, pH=7.4) to a concentration of 0.5˜10 μg / ml, w...

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Abstract

Biomarkers for liver diseases and method for using the same are provided. For detecting liver cirrhosis and liver cancer, the biomarkers are selected from any one of the amino acid sequences with SEQ ID NO:1 to SEQ ID NO:24 or derivatives or fragments or variants or the combination thereof or the antibodies against the amino acid sequences. Then the biomarkers are further developed into detection kits, such that by detecting the existence of autoantibodies or autoantigens in screened specimens, liver diseases are detected with higher accuracy and sensitivity.

Description

BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention is related to biomarkers for liver diseases and method for using the same, in which a method for screening autoantigens is employed to identify biomarkers that can be used in detecting liver diseases. The identified biomarkers are further developed into detection kits to detect the presence of autoantibodies or autoantigens in specimens for screening of liver diseases. [0003] 2. Description of Related Art [0004] People with impaired immune functions are prone to develop immune diseases. The etiology of many human diseases may be traced to our immune system in any of the three conditions described below. The first is reduced immunity, lower activity of immune cells, or reduced quantity of immune cells, such that the human body cannot fight off the invading bacteria, virus or mold, and becomes susceptible to contagious diseases, such as common cold, flu, pneumonia, enteritis, or even hepatitis an...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/47C07K16/18G01N21/78G01N33/543G01N33/53G01N33/567G01N33/574G01N33/68
CPCG01N33/57438G01N2800/085G01N2800/08G01N33/6893
Inventor TSENG, TZU-LINGCHENG, PING-FU
Owner IND TECH RES INST
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