The investigation for high sensitive chemi luminescence method using nanoreactor as a marker

以纳米反应器为标记的高灵敏化学发光方法的研究

• 批准号: 15350038
• 负责人: KAMIDATE Tamio
• 金额: $ 9.41万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15350038/
• 关键词: liposome; nanoreactor; chemi luminescence; peroxidase; ruminol; immunoassay; homogentisic acid γ-lactone; フルオレセイン

Liposomes encapsulated horseradish peroxidase(HRP) were prepared for applying the inner water phase in liposome to the nanoreactor for HRP-catalyzed chemiluminecence(CL) reaction. Phosphatidylcholine(PC), phosphatidylglycerol dimyristoyl(DMPG) and cholesterol were used as a component for liposome. The mole % of PC : DMPG : Chel was 8:1:1. The HRP-trapped liposome was prepared by extrusion method with polycarbonate filter. Homogentisic acid γ-lactone(HAL) and luminol were used as a CL reagent for the determination of horseradish peroxidase (HRP) encapsulated in liposomes. HRP was detected after the lysis of HRP-trapped liposomes with Triton X-100. CL response rate, detection limit and linear range of calibration curve for HRP in HAL CL were compared with those in ρ-iodophenol (ρ-IP) enhanced luminol CL. Maximal light emission in HAL CL appeared more rapidly compared to that in ρ-IP enhanced CL methods, thus resulting in remarkable reduction of CL measurement time. The detection limit for HRP in HAL CL was the same as that in ρ-IP enhanced lnminol CL. The linear range of calibration curve for HRP in HAL CL was improved by factors of 50 compared with that in ρ-IP enhanced luminol CL. From these results, it was found that HAL CL were superior to ρ-IP enhanced luminol CL for the determination of HRP encapsulated in liposomes.

制备了辣根过氧化物酶（HRP）脂质体，并将脂质体内水相应用于纳米反应器中进行HRP催化的化学发光反应。以磷脂酰胆碱（PC）、磷脂酰甘油二肉豆蔻酰（DMPG）和胆固醇为脂质体的组分。PC：DMPG：Chel的摩尔%为8：1：1。采用聚碳酸酯膜挤出法制备包封HRP的脂质体。以高锰酸γ-内酯（HAL）和鲁米诺为化学发光试剂，测定脂质体中辣根过氧化物酶（HRP）的含量。用Triton X-100裂解HRP捕获的脂质体后检测HRP。比较了HRP在HAL和对碘苯酚（ρ-IP）增强鲁米诺化学发光体系中的响应速度、检出限和线性范围。与ρ-IP增强化学发光法相比，HAL化学发光法的最大发光出现得更快，从而显著缩短了化学发光测量时间。HRP在HAL CL中的检出限与ρ-IP增强的lnminol CL相同。与ρ-IP增强的鲁米诺化学发光法相比，HAL化学发光法的线性范围提高了50倍。结果表明，HAL CL法测定脂质体中HRP的灵敏度上级优于ρ-IP增强鲁米诺CL法。

项目成果

リポソームに内封したペルオキシダーゼを触媒に用いるフルオレセイン化学発光による過酸化水素の定量

使用封装在脂质体中的过氧化物酶作为催化剂，通过荧光素化学发光定量过氧化氢

• 期刊: 分析化学 (印刷中)
• 作者: Y.Matsuzaki;A.Nogami;Y.Iwaki;N.Ohta;N.Yoshida;N.Aratani;A.Osuka;K.Tanaka;上舘 民夫
• 通讯作者: 上舘 民夫

Direct detection of horseradish peroxide as a marker molecule encapsulated in liposomes via use of fluorescein chemiluminescence

通过使用荧光素化学发光法直接检测作为封装在脂质体中的标记分子的辣根过氧化物

• 发表时间: 2003
• 期刊: Chemistry Letters 32
• 作者: Y.Ozawa;M.Terashima;M.Mitsumi;K.Toriumi;N.Yasuda;H.Uekusa;Y.Ohashi;Tamio Kamidate
• 通讯作者: Tamio Kamidate

Determination of Peroxidase Encapsulated in Liposomes Using Homogentisic Acid γ-Lactone Chemiluminescence

• DOI: 10.2116/analsci.21.701
• 发表时间: 2005-06
• 期刊: Analytical Sciences
• 影响因子: 1.6
• 作者: T. Kamidate;Nobuhito Kikuchi;A. Ishida;H. Tani

Tamio Kamidate, Yoshiki Ishida, Hirofumi Tani, Akihiko Ishida: "Direct detection of horseradish peroxidase as a marker molecule encapsulated in liposomes via use of fluorescein chemiluminescence"Chemistry Letters. 32・4. 402-403 (2003)

Tamio Kamidate、Yoshiki Ishida、Hirofumi Tani、Akihiko Ishida：“通过使用荧光素化学发光直接检测封装在脂质体中的辣根过氧化物酶”，《化学快报》32·403。

Effects of using a low-copy plasmid and controlling membrane permeability in SOS-based genotoxic bioassay

在基于 SOS 的基因毒性生物测定中使用低拷贝质粒和控制膜通透性的效果

• 发表时间: 2004
• 期刊: Analitica Chimica Acta 522
• 作者: Ogo;S.et al.;Shinobu Kishi;Koji Maehana
• 通讯作者: Koji Maehana