对于石墨电极微阵列的修改,Diazonium-蛋白质添加物:直接和定位的电化学固定
Benjamin P Corgier1, Christophe A Marquette, Loïc J Blum
1Laboratoire de Génie Enzymatique et Biomoléculaire, UMR 5013 EMB2, CNRS Université Claude Bernard Lyon 1, Bât CPE, 43, boulevard du 11 Novembre 1918, 69622 Villeurbanne, Cedex, France.
Journal of the American Chemical Society
|December 22, 2005
概括
这项研究引入了一种新的方法来固定蛋白质,使用直接放置在修改后的蛋白质上的二氧化离子电沉积. 这种技术可以在印电极上精确地对蛋白质进行电位,用于敏感的免疫生物芯片应用.
科学领域:
- 电化学 电化学 电化学
- 表面化学 表面化学
- 生物技术是生物技术.
背景情况:
- 蛋白质固定对于生物传感器的发展至关重要.
- 现有的方法往往缺乏空间特异性和效率.
- 直接电位为受控的蛋白质附着提供了一个潜在的解决方案.
研究的目的:
- 调查用于直接蛋白质固定的二氧化阴离子电位.
- 开发一种电定位方法,将蛋白质附着在印电极上.
- 为了创建一个敏感的免疫生物芯片用于特定抗体检测.
主要方法:
- 用电化学方法将二氧化酸降解到印石墨电极上.
- 免疫球蛋白 (IgG) 与氨酸衍生物的化学合.
- 修饰蛋白质的二亚化以形成aryl diazonium的功能.
- 循环电压测量和X射线光电子光谱学用于表征.
主要成果:
- 在电极表面上成功地对二氧化单层进行了共价接种.
- 在没有交叉通话的情况下,对修改后的子和人类IgG进行电向定位.
- 针对特定抗子IgG检测的免疫生物芯片的演示.
- 对抗子IgG抗体达到50 fmol的低检测极限.
结论:
- 离子电位是直接蛋白质固定化的可行策略.
- 该方法为电气定位生物实体提供了出色的空间特异性.
- 这种新的方法使得高度敏感的免疫生物芯片的开发成为可能.
更多相关视频
相关概念视频
Metal-Ligand Bonds
The hemoglobin in the blood, the chlorophyll in green plants, vitamin B-12, and the catalyst used in the manufacture of polyethylene all contain coordination compounds. Ions of the metals, especially the transition metals, are likely to form complexes.
In these complexes, transition metals form coordinate covalent bonds, a kind of Lewis acid-base interaction in which both of the electrons in the bond are contributed by a donor (Lewis base) to an electron acceptor (Lewis acid). The Lewis acid in...
In these complexes, transition metals form coordinate covalent bonds, a kind of Lewis acid-base interaction in which both of the electrons in the bond are contributed by a donor (Lewis base) to an electron acceptor (Lewis acid). The Lewis acid in...
Diazonium Group Substitution: –OH and –H
Nitrous acid, a weak acid, is prepared in situ via the reaction of sodium nitrite with a strong acid under cold conditions. This nitrous acid prepared in situ reacts with primary arylamines to form arenediazonium salts. Such reactions are known as diazotization reactions. As shown in Figure 1, the formation of arenediazonium salts begins with the decomposition of nitrous acid in an acidic solution to give nitrosonium ions.
1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Mechanism
Nitrous acid is a relatively weak and unstable acid prepared in situ by the reaction of sodium nitrite and cold, dilute hydrochloric acid. In an acidic solution, the nitrous acid undergoes protonation when it loses water to form a nitrosonium ion—an electrophile. Nitrous acid reacts with primary amines to give diazonium salts. The reaction is called diazotization of primary amines.
EDTA: Auxiliary Complexing Reagents
EDTA titrations are usually carried out in highly basic conditions, where the fully deprotonated form of EDTA, Y4−, actively complexes with the free metal ions in the solution. Several metal ions precipitate as hydrous oxide (hydroxides, oxides, or oxyhydroxides) under these conditions, lowering the concentration of free metal ions in the solution. For this reason, auxiliary complexing agents or ligands such as ammonia, tartrate, citrate, or triethanolamine are used in EDTA titrations to...
Masking and Demasking Agents
EDTA titrations may necessitate masking and demasking agents to temporarily protect a particular metal ion in a mixture from the EDTA reaction. These agents facilitate the sequential analysis of the metal ions by forming stable complexes with some—but not all—metal ions during certain steps.
There are many masking agents, such as cyanide, fluoride, triethanolamine, thiourea, and 2,3-bis(sulfanyl)propan-1-ol (formerly 2,3-dimercapto-1-propanol), with the masking agent chosen based on the metal...
There are many masking agents, such as cyanide, fluoride, triethanolamine, thiourea, and 2,3-bis(sulfanyl)propan-1-ol (formerly 2,3-dimercapto-1-propanol), with the masking agent chosen based on the metal...
Complexometric Titration: Ligands
Different monodentate and polydentate ligands are used as complexing agents in complexometric titration reactions. The formation of complexes by mono- and bidentate ligands involves two or more intermediate steps, limiting their use as complexing agents. In comparison, polydentate ligands can form complexes with metal ions in a single-step process, facilitating sharper end points. This means polydentate ligands, such as amino carboxylic acid derivatives, are most commonly employed in...


