The development of nanor eactor for high sensitive chemi luminescence analysis by using liposomes

脂质体高灵敏化学发光分析纳米反应器的研制

• 批准号: 18350037
• 负责人: KAMIDATE Tamio
• 金额: $ 10.39万
• 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: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18350037/
• 关键词: peroxidase; luminol; liposome; chemiluminescence; nanoreactor; ネミノール; ペルオシダーゼ

Horseradish peroxidase (HRP) -encapsulated liposomes were prepared by freeze-thawing method. The number of HRP molecules trapped in liposomes was three times greater than that prepared by extrusion method HRP trapped in liposomes was directly detected by using luminol chemiluminescence (CL) with H_2O_2 without lysis of liposomes. The CL measurement conditions in both a lipid-free bulk solution and in liposomes were optimized in the concentrations of luminol and H_2O_2 by measuring the CL response curves, in which only one peak appeared and the CL intensity was maximal. The CL intensity observed in HRP-catalysed luminol CL in liposomes was a factor of seven greater than that observed in a lipid-free bulk solution. The detection limit in the direct detection of HRP encapsulated in liposomes was sensitive by a factor of 3 compared with that in HRP-catalysed luminol CL in a lipid-free bulk solution. In addition, HRP-trapped liposomes were combined to antibody in order to apply HRP-trapped liposomes to the marker in immunoassay. The CL intensity observed in antibody combined HRP-trapped liposomes was greater 125 times that in antibody combined HRP by avidin-biotin bond.

采用冻融法制备辣根过氧化物酶（HRP）脂质体。脂质体中包封的HRP分子数是挤压法的3倍。用鲁米诺-H_2O_2化学发光法直接检测包封在脂质体中的HRP，不需裂解脂质体。通过测定化学发光响应曲线，优化了在鲁米诺和H_2O_2浓度范围内，无脂质体和脂质体中的化学发光测定条件，使化学发光强度达到最大，且只有一个峰。在脂质体中HRP催化的鲁米诺CL中观察到的CL强度是在无脂质本体溶液中观察到的CL强度的七倍。与HRP催化的鲁米诺CL在无脂质体的本体溶液中相比，直接检测包封在脂质体中的HRP的检测限灵敏度为3倍。此外，将HRP捕获的脂质体与抗体组合，以便将HRP捕获的脂质体应用于免疫测定中的标记物。在抗体结合HRP捕获的脂质体中观察到的CL强度是抗体结合HRP中通过亲和素-生物素键的CL强度的125倍。

项目成果

リン脂質とTriton X-100共存下におけるペルオキシターゼを触媒とするルミノール化学発光の増感効果

磷脂与 Triton X-100 共存时过氧化物酶催化鲁米诺化学发光的增敏作用

• 发表时间: 2008
• 作者: Hiroshi Ohtai;Akihiko Ishida;Hirofumi Tani;and Tamio Kamidate;上舘 民夫
• 通讯作者: 上舘 民夫

Direct determination of horseradish peroxidase encapsulated in liposomes by using luminol chemiluminescence.

使用鲁米诺化学发光法直接测定脂质体中封装的辣根过氧化物酶。

• 发表时间: 2008
• 期刊: Analytical Sciences Vol.24
• 作者: Tamio Kamidate;Kanako Komastu;Hirofumi Tani;and Akihiko Ishida
• 通讯作者: and Akihiko Ishida

Estimation of the membrane permeability of liposomes via use of eosin Y chemiluminescence catalyzed peroxidase encapsulated in liposomes.

通过使用封装在脂质体中的伊红 Y 化学发光催化的过氧化物酶来估计脂质体的膜渗透性。

• 发表时间: 2007
• 期刊: Luminescence Vol.22
• 作者: Tamio Kamidate;Kanako Komastu;Hirofumi Tani;and Akihiko Ishida
• 通讯作者: and Akihiko Ishida

Enhanced luminol chemiluminescence detection of peroxidase in the presence of Triton X- 100 and phospholipid.

在 Triton X-100 和磷脂存在的情况下增强鲁米诺化学发光检测过氧化物酶。

• 发表时间: 2008
• 作者: Hiroshi Ohtai;Akihiko Ishida;Hirofumi Tani;and Tamio Kamidate
• 通讯作者: and Tamio Kamidate

リン脂質とTriton X-100共存下におけるペルオキシターゼを触媒とするルミノール化学発光検出

在磷脂和 Triton X-100 共存中使用过氧化物酶作为催化剂进行鲁米诺化学发光检测

• 发表时间: 2007
• 作者: Hiroshi Ohtai;Akihiko Ishida;Hirofumi Tani;and Tamio Kamidate;上舘 民夫
• 通讯作者: 上舘 民夫