Fabrication of Ag nanoplate-bio tip for evaluation of the binding between proteins and small molecules

用于评估蛋白质和小分子之间结合的Ag纳米板-生物尖端的制造

• 批准号: 23655074
• 负责人: KAWASAKI Hideya
• 金额: $ 2.25万
• 依托单位: Kansai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23655074/
• 关键词: バイオチップ; 銀ナノプレート; 低分子化合物; タンパク質; SALDI-MS; LSPR; LSPRセンサー; 質量分析; バイオチップ; 銀ナノプレート; 低分子化合物; タンパク質; SALDI-MS; LSPR; LSPRセンサー; 質量分析

A complementary analytical methodology combining localised surface plasmon resonance (LSPR) sensing and matrix assisted laser desorption/ionisation-mass spectrometry (MALDI-MS) has been developed by using triangular silver nanoplates (Ag NPLs). Ag NPLs with near-IR LSPR absorbance were prepared via a two-step photo-mediated growth process from Ag nanoparticles. These could be utilised as a common platform for LSPR/MALDI-MS, as they were suitable for LSPR sensing via the analysis of surface plasmon adsorption bands and as the assisting material for LDI-MS. For practical use, the detection of analytes by LSPR sensing can be achieved by using specific biomolecular recognition techniques that involve the surface modification of metal NPs with large molecules such as proteins. We investigated the effect on LSPR sensitivity (i.e. refractive index unit, RIU) of modifying the Ag surface with molecules of different sizes, and found that the RIU values were proportional to the amplitude of the cube root of the molecular weight, Mw1/3. We demonstrated the utility of this complementary LSPR and MALDI-MS analysis methodology by evaluating the binding of soy bean trypsin inhibitor to the Ag NPL substrate covalently modified with trypsin. This specific biomolecule recognition phenomenon was first detected using the LSPR technique, and then successfully identified by the MALDI-MS.

通过使用三角形银纳米板（AG NPLS）开发了一种结合局部表面等离子体共振（LSPR）传感和基质辅助激光解吸/电离 - 质量质谱法（MALDI-MS）的互补分析方法。具有近IR LSPR吸光度的Ag NPL是通过Ag纳米颗粒的两步照片介导的生长过程制备的。这些可以用作LSPR/MALDI-MS的通用平台，因为它们适用于通过表面等离子体的吸附带的分析以及LDI-MS的辅助材料。为了实际使用，可以通过使用特定的生物分子识别技术来实现LSPR传感分析物的检测，该技术涉及具有大分子（例如蛋白质）的金属NP的表面修饰。我们研究了用不同尺寸的分子修饰Ag表面的LSPR敏感性（即折射率单位，RIU）的影响，发现RIU值与分子量量的立方根的振幅成正比，MW1/3。我们通过评估大豆胰蛋白酶抑制剂与用胰蛋白酶共价修饰的Ag NPL底物的结合，证明了这种互补LSPR和MALDI-MS分析方法的实用性。首先使用LSPR技术检测到这种特定的生物分子识别现象，然后由MALDI-MS成功鉴定。