成果報告書詳細
管理番号20110000001164
タイトル*平成22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(4)
公開日2011/9/9
報告書年度2010 - 2010
委託先名国立大学法人東京工業大学
プロジェクト番号P05023
部署名電子・材料・ナノテクノロジー部
和文要約和文要約等以下本編抜粋:1. 研究開発の内容及び成果等 1.高機能性蛍光磁性ビーズの作製と疾患マーカーの高速・高感度検出システムの開発 (1) 1粒子あたり106個以上の蛍光物質を内包する蛍光磁性ビーズの作製 当研究グループでは、磁性酸化鉄のフェライト(ferrite)のナノ粒子をスチレンとグリシジルメタクリレート(glycidiyl methacrylate,GMA)の共重合体で被覆し、さらに最表面をポリGMAで被覆した粒径200nmのアフィニティ磁性ビーズ(FGビーズ)を開発してきた(図1)。
英文要約1. Preparation of highly functionalized fluorescent magnetic beads and development of high-speed and high-sensitive disease-related marker detection system
(1) Preparation of novel fluorescent magnetic beads with over 106 fluorophores per one bead
We have developed 200 nm affinity magnetic beads (FG beads), which several ferrite nanoparticles are coated by polymers including poly(glycidyl methacrylate (GMA)). In FY 2009, we achieved preparation of novel fluorescent magnetic beads which both ferrite nanoparticles and fluorophores are encapsulated into polymers. As fluorophores, we selected hydrophobic Eu(TTA)3(TOPO)2 (TTA: thenolytrifluoroacetone, TOPO: tri-n-octylphosphine oxide). Naming after fluorescence and ferrite, we labeled the beads as “FF beads”. We also succeeded in preparation of more magnetically responsive 140 nm FF beads. In FY 2010, we examined high-speed and high-sensitive disease-related marker detection system using 140 nm FF beads.
(2) Preparation of highly functionalized FF beads on which an antibody is fixed
In FY 2009, we established fixation method of antibodies recognizing a disease-related marker on FF beads in buffer through amide bonding between amines in the antibodies and carboxylic acids on FF beads. In FY 2010, we examined conditions that the antibodies were fixed on epoxy FF beads talking advantage of imidazoles in the antibodies and found out that Fab fragments and/or F(ab’)2 fragment antibodies also could be fixed on FF beads.
(3) Construction of magnetic collection technology of highly functionalized FF beads by means of external magnetic field such as magnet
We examined magnetic collection time of FF beads for sandwich immunoassay to detect antigens and found out that only 1 minute magnetic collection of FF beads was enough for highly sensitive detection of antigens.
(4) High-speed, high-efficient, and high-sensitive detection of disease-related markers taking advantage of magnetic collection of FF beads on which antibodies recognizing the disease-related markers are fixed
Since we found that FF beads on which an antibody recognizing an antigen are fixed can be collected by magnet to a specific place, we examined detection of disease-related markers taking advantage of magnetic collection of FF beads. In FY 2009, we examined high-speed, high-efficient, and high-sensitive detection of BNP (brain natriuretic peptide, a marker of heart disease) using FF beads on which the antibody recognizing BNP is fixed, taking advantage of magnetic collection, and could detect 2.0 pg/ml of BNP for only 5 minutes sandwich immunoassay. In FY 2010, we investigated detection of PSA (prostate specific antigen, a marker of prostate cancer) using FF beads and found out that 0.1 ng/ml of PSA could be detected within 5 minutes sandwich immunoassay with only 1 minute magnetic collection of anti-PSA antibody-fixed FF beads. In addition, sandwich immunoassay using F(ab’)2 fragment anti-PSA antibody-fixed FF beads enabled us to detect 0.01 ng/ml of PSA. Based on these findings, we attempted high-speed and high-sensitive detection of PSA in serum and could achieve sensitive detection of PSA. Thus, we concluded that the developed system detecting disease-related markers taking advantage of magnetic collection of FF beads had a high correlation with conventional immunoassays.
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