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A marker comprising anti-CK8/18 complex autoantibody and its use for diagnosing cancer

一种自身抗体、复合物的技术，应用在抗动物/人类的免疫球蛋白、抗受体/细胞表面抗原/细胞表面决定因子免疫球蛋白、引入外来遗传物质而修饰的细胞等方向，能够解决诊断的生物标记物局限、诊断效果不令人满意、识别诊断标记物困难等问题

Active Publication Date: 2013-12-04
KOREA RES INST OF BIOSCIENCE & BIOTECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although previous studies on autoantibodies have shown that autoantibodies can be used as cancer-associated markers, their diagnostic performance is unsatisfactory and there are limitations in the use of autoantibodies as biomarkers for cancer diagnosis
In addition, since the autoantibody detection method does not include a large number of medical records or actually requires too many tests, the autoantibody detection method has limitations in practical application
Indeed, difficulties remain in identifying autoantibodies as diagnostic markers for cancer

Method used

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  • A marker comprising anti-CK8/18 complex autoantibody and its use for diagnosing cancer
  • A marker comprising anti-CK8/18 complex autoantibody and its use for diagnosing cancer
  • A marker comprising anti-CK8/18 complex autoantibody and its use for diagnosing cancer

Examples

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example 13

[0050] Analytical methods used to measure expression levels include, but are not limited to, western blot, ELISA (enzyme-linked immunosorbent assay), radioimmunoassay (otA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, immunohistochemical staining, immunoassay Precipitation analysis, complement fixation assay, FACS, and protein chip analysis. Through the above detection method, the formation of antigen-antibody complexes in normal control samples can be compared with the formation of antigen-antibody complexes in individuals with breast cancer or suspected of having breast cancer, thereby diagnosing suspected breast cancer. Incidence of breast cancer in patients with breast cancer.

[0051] The method for diagnosing breast cancer can be realized by the antibody-antigen reaction between the cytokeratin 8 / 18 complex-specific autoantibody of the present invention and an antigen specifically binding to the autoantibody. As used herein, the term...

Embodiment 1

[0081] Example 1. Acquisition of cell populations producing autoantibodies and cells producing K94 autoantibodies

[0082] In order to acquire autoantibodies produced during carcinogenesis, H-ras12V transgenic mice reported to form liver cancer similar to human liver cancer were used. Splenocytes were obtained from H-ras12V transgenic mice with liver cancer as a B cell panel, and the splenocytes were fused with mouse myeloma cells Sp2 / 0 to prepare a B cell hybridoma cell line. Perform cell fusion according to common B-cell hybridoma preparation methods. Primary selection of fusion cells was performed using HAT medium (hypoxanthine-aminopterin-thymidine medium), and only clonogenic cells were isolated and cultured. Of these cultured cells, only cells in the culture medium for which antibodies reactive with cancer cells were detected were selected and maintained.

[0083] Autoantibody reactivity to cancer cells was examined by flow cytometric analysis of cancer cell lines af...

Embodiment 2

[0086] Example 2. Purification of TAB-K94 monoclonal antibody

[0087] For the analysis of antigens against TAB-K94 autoantibodies, purified antibodies are required. A cell culture medium obtained by culturing a large number of TAB-K94 antibody-producing clones or TAB-K94 antibody-producing cells was injected into the peritoneal cavity of a mouse to obtain ascites fluid, which was used for antibody purification. The type of TAB-K94 antibody was identified as IgM using a commonly used isotype ELISA. MBP-agarose (mannose-binding protein-immobilized sepharose: Pierce) or protein L sepharose was used in affinity chromatography for IgM purification. After SDS-electrophoresis, purified antibodies were checked by Coomassie staining and protein quantification was performed before use ( figure 2 ).

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Abstract

The present invention relates to a cytokeratin 8 / 18 complex-specific autoantibody or a fragment comprising an antigen-binding site (paratope) thereof, the use thereof in the diagnosis of breast cancer, a polypeptide having an amino acid sequence of an epitope specifically binding to the autoantibody, a composition for diagnosing breast cancer comprising an agent capable of measuring an expression level of the autoantibody or the fragment comprising an antigen-binding site thereof, a hybridoma cell line producing the autoantibody, and a kit for diagnosing breast cancer comprising the composition of the present invention. Further, the present invention relates to a method for diagnosing breast cancer, comprising the step of detecting the cytokeratin 8 / 18 complex-specific autoantibody or the fragment comprising the antigen-binding site thereof using the composition of the present invention, and a method for screening a therapeutic agent for breast cancer using the autoantibody.

Description

technical field [0001] The present invention relates to: a cytokeratin 8 / 18 complex-specific autoantibody or its fragment containing an antigen-binding site (paratope); the use of this autoantibody in the diagnosis of breast cancer; A polypeptide of the amino acid sequence of an epitope that is sexually bound; a composition for diagnosing breast cancer, the composition comprising the ability to measure the expression level of the autoantibody or the expression level of a fragment of the autoantibody comprising an antigen-binding site a reagent; a hybridoma cell line producing the autoantibody; and a kit for diagnosing breast cancer comprising the composition of the present invention. Furthermore, the present invention relates to a method for diagnosing breast cancer, the method comprising the steps of: using the composition of the present invention to detect cytokeratin 8 / 18 complex-specific autoantibodies or detecting autoantibodies comprising antigen binding site; and the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N5/16G01N33/574
CPCG01N2333/4742C07K7/06G01N33/57415C07K16/18G01N33/574C07K16/30C07K2317/32C07K2317/34C12N5/16
Inventor 赵恩伟许昌圭高正宪禹美京俞香淑黄海民
Owner KOREA RES INST OF BIOSCIENCE & BIOTECHNOLOGY
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