Characteristics of a liposome immunoassay on a poly(methyl methacrylate) surface

被引:21
|
作者
Hwang, Sang Youn
Kumada, Yoichi
Seong, Gi Hoon
Choo, Jaebum
Katoh, Shigeo
Lee, Eun Kyu [1 ]
机构
[1] Hanyang Univ, Dept Chem Engn, Ansan, South Korea
[2] Kyoto Inst Technol, Dept Chem & Mat Technol, Kyoto 606, Japan
[3] Hanyang Univ, Dept Appl Chem, Ansan, South Korea
[4] Kobe Univ, Dept Sci & Chem Engn, Kobe 657, Japan
基金
日本学术振兴会;
关键词
liposome immunoassay; immunoliposome; PMMA; microarray detection; signal enhancement;
D O I
10.1007/s00216-007-1614-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Liposome immunoassay (LIA) is based on enzyme immunoassay (EIA) but the detection sensitivity could be significantly enhanced by using antibody-coupled immunoliposomes encapsulating HRP (horse radish peroxidase). Here, we applied LIA to non-porous poly(methyl methacrylate) (PMMA) and polystyrene (PS) surfaces to compare its detection sensitivity with that of EIA, using rabbit IgG (a ligand molecule) and anti-rabbit IgG antibody (a capture molecule) as the model system. LIA developed much stronger color signals than EIA, especially at a lower concentration range (< ca. 1 mu g mL(-1)). PMMA showed higher affinity toward rabbit IgG than the PS surface, and the anti-rabbit IgG antibody adsorbed on PMMA was more stable than that on PS. Furthermore, the effects of spot volume and antibody concentration on the signal density were analyzed. The signal density increased as the antibody concentration increased, but it was not significantly affected by the spot volume (2.5-20 mu L). In conclusion, LIA on PMMA as a solid support is a very useful, highly sensitive microarray detection system.
引用
收藏
页码:2251 / 2257
页数:7
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