Immobilisation of multilayer bioreceptor assemblies on solid substrates.
Immobilisation of multilayer bioreceptor assemblies on solid substrates.
复制标题
将多层生物受体组件固定在固体基质上。
DOI：
10.1016/s0956-5663(97)00107-3
复制
发表时间：
1998
影响因子：
12.6
通讯作者：
J. Ramsden
中科院分区：
文献类型：
--
作者：
E. Brynda;M. Houška;J. Škvor;J. Ramsden
多层组件的制备通过交替吸附的单克隆抗体对辣根过氧化物酶（抗HRP）和硫酸葡聚糖（DS）在酸性pH值的固体支持物上的单层。交联后，用戊二醛，DS被洗出的膜与缓冲生理盐水，而抗体保持固定在支持物上。通过红外多内反射光谱法在锗载体上原位监测组装。通过ELISA和通过光波导光模式光谱法测量固定化抗体对HRP的结合能力。固定化的抗HRP双层的活性是通过简单的物理吸附制备的单层的约两倍。添加另外的抗HRP层可以将活性仅增加至单层活性的2·5，而与组装体中的层数无关。从人血浆中的蛋白质的非特异性吸附是三倍低的固定化的抗HRP多层膜上的表面上只覆盖有物理吸附的抗HRP单层。
Multilayer assemblies were prepared by alternating adsorption of monolayers of monoclonal antibody against horse radish peroxidase (anti-HRP) and dextran sulfate (DS) on solid supports at acid pH. After crosslinking with glutaraldehyde, DS was washed out of the film with buffered physiological saline, while the antibody remained immobilised on the support. Assembly was monitored in situ on germanium supports by infrared multi-internal reflection spectroscopy. The binding capacity of the immobilised antibodies for HRP was measured by ELISA and by optical waveguide light mode spectroscopy. The activity of an immobilised anti-HRP bilayer was approximately twice that of a monolayer prepared by simple physiosorption. An addition of further anti-HRP layers could increase the activity only up to 2·5 of the monolayer activity independently of a number of layers in the assembly. The non-specific adsorption of proteins from human blood plasma was three times lower on the immobilised anti-HRP multilayer film than on the surface covered only with a physisorbed anti-HRP monolayer.