成果報告書詳細
管理番号20100000001588
タイトル*平成21年度中間年報 ナノテク・先端部材実用化研究開発/高機能性蛍光磁性ビーズによる高速・高感度疾患診断システムの開発
公開日2011/1/18
報告書年度2009 - 2009
委託先名学校法人慶應義塾
プロジェクト番号P05023
部署名ナノテクノロジー・材料技術開発部
和文要約和文要約等以下本編抜粋:1.研究開発の実績 1.高速・高感度疾患診断に利用する疾患マーカーの検証 (1)腫瘍疾患マーカーの検証 慶応義塾大学のグループでは既に有しているEGFRに対する抗体を産生する3種類(225,528、B4G7)とc-erbB2産物については2種類(NB3,TA1)のハイブリドーマーから抗体を精製した。抗体の特徴は図1-1、図1-2に記す。
すなわち、今年度はEGFRとc-erbB2産物に対する抗体を産生するハイブリドーマーをヌードマウスの腹腔内に移植し腹水を作製し採取し、以下の図2のような工程を経て腹水からアフィニティークロマトを用いてそれぞれの抗体を精製した。
英文要約Department of Surgery, Keio University, School of Medicine
Kitagawa MD,PhD
Title:Development of high speed and sensitive diagnostic systems for the disease by highly functionalized ferrite fluorescent beads (FY2009-FY2010) FY2009 Annual Report
Summary:The over-expression of epidermal growth factor receptors (EGFR) in squamous cell carcinoma and c-erbB2 products in adnocarcinoma was observed in these cancers. As the diagnosis of these cancer patients was easily performed by the detection of these products in there bloods, the high speed and sensitive diagnostic systems is required by clinics.
To develop high speed and sensitive diagnostic systems for squamous cell carcinoma and adnocarcinoma using highly functionalized ferrite fluorescencent beads (FF beads), we purified anti-epidermal growth factor receptor (EGFR) and anti-c-erbB2 product antibodies. Three kinds of hybridomas (225,528,B4G7) for EGFR and two kinds of hybridomas (NB3,TA-1) for c-webB2 were implanted in the abdominal cavity of nude mice and the ascites was collected from the nude mice.. The antibodies were purified from ascites by MabTrap Kit (GE). These antibodies were immobilized onto the surfaces of the FF beads and succeeded in accelerating specific antigen-antibody reactions when FF beads fixed with antibodies were magnetically collected onto complementary reaction regions on solid substrates.
To evaluate the clinical use of these assay systems using FF beads, the clinical samples should be assessed. The protocol to collect the blood samples from the patients was approved by the ethics committee of Keio University, school of medicine. Written informed consent was obtained form each patient prior to study entry. All samples were immediately frozen at -80℃. We collected more than 30 samples from the cancer patients in this year.
The information of tumor markers and its antibodies for squamous cell carcinoma and adenocarcinoma in human was collected and analyzed to establishment of the new assay systems.
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