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Marker for diagnosing liver cancer containing anti-ATIC autoantibody, and composition for diagnosing liver cancer containing antigen thereof

An autoimmune antibody and composition technology, applied in the field of cleotide formyltransfera, can solve the problems of limited implementation, low practicability, and little diagnostic effect, and achieve effective development results

Active Publication Date: 2014-04-16
KOREA RES INST OF BIOSCI & BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is a problem that it can be detected when the antigenic determinant is an ordinal epitope depending on the protein sequence, but cannot be detected when it is a hypothetical epitope depending on the protein structure
[0010] As mentioned above, studies on autoimmune antibodies that can be used as existing markers related to cancer occurrence are not very practical and have little diagnostic effect. Therefore, it is still difficult to use autoimmune antibodies as biomarkers for cancer diagnosis. there are certain problems
Moreover, the detection method of autoimmune antibodies cannot cover so many situations, and requires too many experiments, and its implementation is still limited, so it is still impossible to discover the markers of autoimmune antibodies for the diagnosis of liver cancer

Method used

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  • Marker for diagnosing liver cancer containing anti-ATIC autoantibody, and composition for diagnosing liver cancer containing antigen thereof
  • Marker for diagnosing liver cancer containing anti-ATIC autoantibody, and composition for diagnosing liver cancer containing antigen thereof
  • Marker for diagnosing liver cancer containing anti-ATIC autoantibody, and composition for diagnosing liver cancer containing antigen thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1: Obtaining XC154 autoimmune antibodies derived from HBx mice

[0083] In order to obtain autoimmune antibodies produced together with cancer, HBx-transformed mice were used, and it is known that the liver cancer that develops therein has a morphology similar to that of human liver cancer. From HBx-transformed mice in breeding, the spleen (spleen) cells of individuals whose liver cancer was confirmed were taken as B cells, and cell fusion was performed with mouse myeloma cells Sp2 / 0 to prepare B-cell hybridomas cell. Cell fusion was performed according to the general method for preparing B-cell hybridoma cells. The fused cells were screened once using hypoxanthine-aminopterin-thymidine medium (HAT medium), and only cells that formed clones were cultured. In the cell culture medium, the cells that can detect the cancer cell reaction antibody are selected again, and the culture is continued.

[0084] The reactivity of autoimmune antibodies to cancer cells was ...

Embodiment 2

[0086] Example 2: Analysis of the Complementary Determining Region (CDR) of the XC154 antibody and purification of the antibody

[0087] After confirming that the XC154 antibody can specifically recognize the antigen expressed in cancer cells, in order to obtain information on the antigen recognition specificity of the XC154 antibody, the complementarity determining region sequences of the antibody were analyzed. The detailed method is as follows:

[0088] collect 10 6 About XC154 antibody-producing cells were used to extract total RNA using an RNA extraction kit (Qiagen). Complementary DNA synthesis (complementary DNA (cDNA) synthesis) kit (Invitrogen) was used to synthesize cDNA from 5 μg of total RNA, and the synthesized 1 μg of cDNA and mouse heavy chain constant region primers (Mouse Heavy chain constant region primer) F5'-CTT CCG GAA TTC S AR GTN MAG CTG SAG SAG TCW GG-3' (SEQ NO.21), R5'-GGA AGA TCT GAC ATT TGG GAA GGA CTG ACT CTC-3' (SEQ NO .22) and mouse light chai...

Embodiment 3

[0090] Example 3: Confirmation of the expression of XC154 antibody-specific reactive antigens in various cancer cells

[0091] In order to confirm the reactivity of the XC154 antibody to various cancer cell lines, various cancer cells were stained intracellularly by the method of Example 1 above, and analyzed by flow cytometry ( image 3 ). From the results, it was confirmed that the XC154 antibody-reactive protein was overexpressed in liver cancer cell lines such as HepG2 and SK-Hep-1, and various types of cancer cell lines such as HeLa, LNcap-LN3, and A549.

[0092] In addition, in order to confirm the expression position in the cell, intracellular staining was performed and observed with a confocal fluorescence microscope, and relatively strong staining was confirmed at the position of the cytoplasm or the position of the cell membrane ( image 3 ).

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Abstract

The present invention relates to a fragment comprising an autoantibody which specifically binds to ATIC or an antigen-binding site (i.e. paratope) of the autoantibody, a composition for diagnosing liver cancer containing an agent which measures the expression level of the fragment, a hybridoma cell line producing the autoantibody, a diagnostic kit for liver cancer containing the composition, a method for diagnosing liver using the composition, and a method for screening a liver cancer therapeutic agent using the autoantibody. Using the anti-ATIC specific autoantibody of the present invention as a marker for diagnosing liver cancer, the occurrence of liver cancer can be diagnosed using a non-invasive biological sample such as blood, plasma, serum, lymph tissue, and the like, with a sensitivity of about 87% and a specificity of about 88%, and as liver cancer can be easily diagnosed using an amino acid sequence identified in the present invention, the autoantibody is effective for developing a diagnostic kit for liver cancer.

Description

technical field [0001] The present invention relates to a self-specific combination with ATIC (5-aminoimidazole-4-carboxamide ribon ucleotide formyltransferase / IMP cyclohydrolase, 5-aminoimidazole-4-carboxamide nucleotide formyltransferase / IMP cyclohydrolase) Immune antibody (auto antibody) or a fragment containing its antigen-binding site (antigen-binding site, Paratope); an antigen fragment that specifically binds to the autoimmune antibody; A composition for diagnosis of liver cancer according to the expression level of fragments of the antigen-combining site; a hybridoma cell line producing the autoimmune antibody and a kit for diagnosis of liver cancer comprising the composition of the present invention. Furthermore, the present invention relates to a method for diagnosing liver cancer using the composition of the present invention and a method for screening a therapeutic agent for liver cancer using the autoimmune antibody. Background technique [0002] In South Korea...

Claims

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Application Information

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IPC IPC(8): C07K16/30G01N33/53G01N33/574C12N5/16
CPCC07K16/40G01N33/564G01N33/57438C07K16/30C12N5/16G01N33/53G01N33/574
Inventor 赵恩伟许昌圭高正宪禹美京兪香淑黃海民
Owner KOREA RES INST OF BIOSCI & BIOTECH
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