Study of the planar membrane on the silica compounds and its application

二氧化硅化合物平面膜的研究及其应用

• 批准号: 61880024
• 负责人: NAKANISHI Mamoru
• 金额: $ 2.62万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61880024/
• 关键词: Lipid hapten; Planar membrane; FET; Antibody; LB膜; ラングミュアブロジェット法; ナノ秒蛍光; スペーサー; ラングミュア・ブロジェット; 免疫センサー; マクロファージ; 蛍光エネルギー移動

We have studied the antibody-dependent binding of macrophages to supported planar lipid monolayers containing haptented phospholipids. The binding of macrophages to planar DMPC or DPPC monolayers was dependent on IgG subclasses and hapten concentrations. The binding efficiencies were as follows on both 'solid' and 'fluid' lipid monolayer membranes; IgGl IgG2a IgG2b IgG3. Next, we determined the location of haptens (TNP and dansyl) in lipid bilayer membranes by nanosecond fluorometry. It was found that TNP (or dansyl) residues of lipid haptens locate at the similar vertical position on the membrane surfaces even if they have different length spacers. From these kinds of experiments it was explained how the spacer length can modulate the binding affinity of antibody molecules to the lipid hapten in the membrane. Lastly, we have studied a new type of immuno-FET which has lipid membranes on the surface of an ISFET gate (SI_3N_4/SiO_2) by the fusion of single unilamella liposomes containing lipid haptens (TNP-Cap-DPPE). When mouse monoclonal anti-TNP IgGl was added to the solution, a time-dependent potential change (5-10mV) towards negative was odsedved by a pH/pCO_2 monitor. As for a control experiment, non-specific muse IgGl was added to the solution, no potential change was observed.

我们研究了巨噬细胞与含有半抗原磷脂的支撑平面脂质单层的抗体依赖性结合。巨噬细胞与平面 DMPC 或 DPPC 单层的结合取决于 IgG 亚类和半抗原浓度。 “固体”和“液体”脂质单层膜上的结合效率如下； IgGl IgG2a IgG2b IgG3。接下来，我们通过纳秒荧光测定法确定了半抗原（TNP 和丹磺酰）在脂质双层膜中的位置。研究发现，脂质半抗原的 TNP（或丹磺酰）残基位于膜表面相似的垂直位置，即使它们具有不同长度的间隔基。从这些类型的实验中，解释了间隔基长度如何调节抗体分子与膜中脂质半抗原的结合亲和力。最后，我们研究了一种新型免疫FET，它通过融合含有脂质半抗原的单层脂质体（TNP-Cap-DPPE）在ISFET栅极（SI_3N_4/SiO_2）表面具有脂质膜。当将小鼠单克隆抗TNP IgG1添加到溶液中时，通过pH/pCO_2监测器观察到朝向负值的时间依赖性电位变化(5-10mV)。至于对照实验，将非特异性muse IgG1添加到溶液中，没有观察到潜在的变化。

项目成果

中西守: 化学と薬学の教室(バイオテクノロジー特集). 81-85 (1988)

Mamoru Nakanishi：化学与药学课堂（生物技术专题）81-85（1988）。

Kato,K.; Nakanishi,M.; Arata,Y.; Miyamoto,H.: "Calcium Influx in a Single Rat Basophilic Leukemia Cell as Revealed with a Digital Imaging Fluorescence Microscope." J. Biochem.102. 1-4 (1987)

加藤，K.；

中西守: "生命科学の基礎6.(抗原認識機講)" 学会出版センター, (1989)

中西守：《生命科学基础6.（抗原识别机制）》学会出版中心，（1989）

中西守: 代謝. 24. 209-215 (1987)

中西守：新陈代谢。24. 209-215 (1987)

Mizuguchi,J.; Utsunomiya,N.; Nakanishi,M.; Arata,Y.: "Phorbol Myristate Acetate Inhibits Increase in Membrane Fluidity Induced by Anti-IgM in B Cells." J. Immunology. 140. 2495-2499 (1988)

水口，J.；