Identifications of diagnostic biomarkers specific binding to soluble proteins of adenocarcinoma A 549 cell lines by an autoantibodiomics
| Project/Area Number |
17590501
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Osaka Medical College |
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Principal Investigator |
NAKANISHI Toyofumi Osaka Medical College, Faculty of Medicine, Associate Professor, 医学部, 助教授 (10247843)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Akira Osaka Medical College, Faculty of Medicine, Emeritus Professor, 医学部, 名誉教授 (00028581)
TAKEUCHI Toru Osaka Medical College, Faculty of Medicine, Research Associate, 医学部, 助手 (10330078)
MURAO Hitoshi Osaka Medical College, Faculty of Medicine, Research Associate, 医学部, 助手 (00278540)
GOTO Isao Osaka Medical College, Faculty of Medicine, Assistant Professor, 医学部, 講師 (90330084)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Autoantibody / Cancer / Proteomics / Adenocarcinoma / Biomarker / Diagnosis / 癌プロテオミクス / 自己抗体 / 肺アデノカルシノーマ / 肺癌バイオマーカー / 2D-PAGE / α-エノラーゼ / ペルオキシレドキシン-6 |
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| Research Abstract |
At 2001, Prof. Hanash and his colleagues first reported that the autoantibodies against Annexin-I ＆ II frequently (37-40%) were existed in plasma of lung adenocarcinoma using proteomics-based analysis. Thereafter many researchers have been identified autoantibody in cancer patients' plasma against targeted proteins derived from cancer cells using the same technique. We also applied the novel proteomics-based analysis to identify the candidate biomarkers for the diagnosis of lung adenocarcinoma. The advantages of the identification of autoantibodies against cancer/tumor related antigens were more easy and simple than the gene analytical method. This technique, so-called autoantibodiomics, was expected to apply the screening of cancer diagnostic biomarkers in plasma. In this project we identified candidate new lung cancer diagnosis markers.
[Method] We treated a cultured human lung cancer cell strain (A549) in solubilization buffer solution and soluble fractions were applied a sample solution. The soluble proteins were separated with a two-dimensional electrophoresis and then transferred on a PVDF membrane. After blocking the PVDF membrane, plasma (500 times dilution) as a primary antibody was reacted with separated proteins on membrane, detected a positive spots using ECLplus reagent Kit. On the other hand, silver-staining gel was cut-off a positive spots corresponding to the ECL positive spots and analyzed them by a time-of-flight mass spectrometer (UltraFlex) or an ion trap type mass spectrometer ( LCQDECA) and a database searched a provided result and identified target proteins.
[Results and conclusion] We found eight kinds of biomarkers, anti α-enlace, chaperonin, peroxiredoxin-6 autoantibodies, in patients' plasma of lung adenocarcinoma. We concluded that these autoantibodies were candidate biomarker to diagnose lung adenocarcinoma.
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Report
(3 results)
Research Products
(7 results)
