Direct oriented immobilization of F(ab′) antibody fragments on gold

被引:72
作者
Brogan, KL [1 ]
Wolfe, KN [1 ]
Jones, PA [1 ]
Schoenfisch, MH [1 ]
机构
[1] Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA
关键词
antibody immobilization; immunoassay; quartz crystal microbalance; F(ab ') antibody fragments;
D O I
10.1016/S0003-2670(03)00991-7
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The preparation of oriented immunosurfaces by the direct immobilization of F(ab') fragments of rabbit anti-calf alkaline phosphatase (RACAP) Immunoglobulin G (IgG) onto gold (An) surfaces via the formation of a Au-thiolate bond is described. In order to compare specific and random immobilization methods, F(ab')-modified An is compared to immunosurfaces of non-specifically adsorbed F(ab) fragments of RACAP IgG. The immobilization chemistries of the resulting antibody-modified Au surfaces are characterized with X-ray photoelectron spectroscopy (XPS). Antigen binding is characterized using both a quartz crystal microbalance (QCM) and a solid phase enzyme linked immunosorbent assay (ELISA). We demonstrate that specifically immobilized F(ab') fragments can be used to produce immunosurfaces with higher antigen-binding efficiencies than corresponding surfaces modified with randomly immobilized antibody fragments. The relationship between antigen-binding efficiency and the surface coverage of immobilized antibody fragments is also discussed. (C) 2003 Published by Elsevier B.V.
引用
收藏
页码:73 / 80
页数:8
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