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New applications of antibody of GDF15 (Growth differentiation factor 15) protein

A protein and antibody technology, applied in the direction of anti-growth factor immunoglobulin, anti-animal/human immunoglobulin, microbial determination/examination, etc.

Active Publication Date: 2010-10-06
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the studies are on the correlation between GDF15 and tumors and cardiovascular diseases, and there is no report on the correlation between viral hepatitis and GDF15

Method used

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  • New applications of antibody of GDF15 (Growth differentiation factor 15) protein
  • New applications of antibody of GDF15 (Growth differentiation factor 15) protein
  • New applications of antibody of GDF15 (Growth differentiation factor 15) protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Discovery of hepatitis virus infection-induced cytokine GDF15

[0030] Huh7 cells (purchased from Apath, USA) were infected with clinically isolated acute phase 2a genotype HCV, and the cells were collected at different time points for gene chip determination (gene chips were purchased from Boao Biological Co., Ltd. and biochip Beijing National Engineering Research Center, the product is a 35K human mRNA oligonucleotide probe chip, the specific chip operation is completed by Boao Biotech according to its standard operating procedures), and the results screened out GDF15, one of the highly expressed genes after HCV infection, which was obtained from From the second day after infection, it was significantly increased by more than ten times. Therefore, it may play a certain role in the regulation of HCV infection. For specific results, see figure 1 .

Embodiment 2

[0031] Example 2, Cytokine GDF15 expression level and mutual regulation of HCV replication

[0032] 1. At the cellular level, HCV infection can promote the upregulation of GDF15 expression

[0033] Huh7 cells were infected with HCV virus isolated from the clinical acute phase of hepatitis C, and the cells and cell supernatant were collected at different time points; real-time fluorescent quantitative (Real-time PCR) method was used to detect the relative quantification of GDF15 mRNA in the cells, and the QIAGEN company The one-step RT-PCR quantitative kit (article number: 204154) and the 7900 real-time fluorescent quantitative PCR instrument of ABI Company were detected according to the standard procedure recommended by the manufacturer, and the amplification primer sequence was as follows: Upstream primer: 5`-ACT TGT TAG CCA AAG ACT GCC-3`, downstream primer: 5`-AAC CTT GAGCCC ATT CCA-3`. see results figure 2 , HCV-infected cells in vitro can significantly induce the trans...

Embodiment 3

[0037] Example 3, Regulatory Mechanism of GDF15 Expression Stimulated by HCV Infection

[0038] Huh7 cells were transfected with 3 structural protein genes and 5 nonstructural protein genes of HCV, and the supernatant was collected after 48 hours. The protein level of GDF15 was detected by ELISA. Compared with the empty vector negative control, the structural proteins core, E1 and E2 Can significantly induce the up-regulation of GDF15 expression, but the non-structural proteins P7, NS2, NS3, NS3 / 4A and NS5B did not induce the increase of GDF15 expression level, the results are shown in Figure 5 . The structural genes core, E1 and E2 of HCV up-regulate GDF15, which proves that the regulatory effect of cytokine GDF15 on HCV may be related to the combination of GDF15 and HCV structural protein.

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Abstract

The invention discloses new applications of an antibody of GDF15 (Growth differentiation factor 15) protein. The invention provides the following three applications of the antibody of the GDF15 protein: 1. the application in preparing a reagent for assisting to identify a hepatitis C patient or a hepatitis B patient; 2. the application in preparing a reagent kit for assisting to identify the hepatitis C patient or the hepatitis B patient; and 3. the application in preparing a reagent kit for assisting to identify the hepatitis C progress of the hepatitis C patient, wherein the hepatitis C progress is chronic hepatitis C, hepatitis C cirrhosis or liver cancer. On the basis of the three applications, the antibody of the GDF15 protein can be prepared into three reagent kits. The reagent kit can be used for the serological diagnosis of viral hepatitis (the hepatitis C or the hepatitis B) and judging the disease progress (the chronic hepatitis C, the hepatitis C cirrhosis or the liver cancer) and for prognosis and viral hepatitis treatment monitoring and has great value for the diagnosis and the treatment of the hepatitis C and the hepatitis B.

Description

technical field [0001] The present invention relates to a new application of the antibody of GDF15 protein. Background technique [0002] Hepatitis C virus (HCV) is a single positive-sense RNA virus belonging to the Flaviviridae family, and is an important pathogenic factor of acute and chronic hepatitis transmitted by blood. The chronic rate of HCV infection is very high, ranging from 50% to 85%. Chronic HCV infection is closely related to liver cirrhosis and hepatocellular carcinoma, which seriously endangers people's health. At present, there are nearly 200 million HCV infected people in the world, and the infection rate is 3.1% (Lauer GM, Walker BD. Hepatitis C virus infection. N Engl J Med. 2001 Jul 5; 345(1): 41-52.). my country is one of the high-incidence areas of hepatitis C. The infection rate of the general population in my country is 3.2%, and the number of infected people is about 42 million (Chinese Medical Association, "Guidelines for the Prevention and Treat...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/577G01N33/574C12Q1/68C07K16/22
Inventor 杨威刘秀英司有辉
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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