Stepwise silver-staining-based immunosorbent assay for amyloid-β autoantibody detection

被引：5

作者：

Shim, So-Youn ^{[1
]}

Woo, Jung-Reem ^{[1
]}

Nam, Eun-Ju ^{[2
]}

Hong, Hyun-Joo ^{[3
]}

Mook-Jung, Inhee ^{[3
]}

Kim, Young-Ho ^{[2
]}

Nam, Jwa-Min ^{[1
]}

机构：

[1] Seoul Natl Univ, Dept Chem, Seoul, South Korea

[2] Digital Bio Tech, Seoul, South Korea

[3] Seoul Natl Univ, Coll Med, Dept Biochem & Canc Res, Seoul, South Korea

来源：

NANOMEDICINE | 2008年 / 3卷 / 04期

关键词：

Alzheimer's disease; amyloid-beta autoantibody; gold nanoparticle; immunosorbent assay; silver staining;

D O I：

10.2217/17435889.3.4.485

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Aims: To detect amyloid-beta (A beta) autoantibodies in a reliable and high-throughput fashion, we developed a stepwise silver-staining-based immunosorbent assay in a 96-well-plate platform. Materials & methods: A beta autoantibodies were incubated in an A beta-immobilized 96-well microplate. Antihuman IgG-modified gold nanoparticle probes were then used to bind to the autoantibodies and signal enhancement was carried out with stepwise silver-staining on immobilized gold nanoparticle probes. A microplate reader was used to quantify silver-stained gold nanoparticles on a well-plate surface. Results & discussion: Stepwise silver-staining at low temperature enables long-term silver-staining with minimal increase of background signal. This stepwise staining method helps solve the problems of one-step staining, such as nonspecific binding or nonuniformity in silver precipitation after prolonged silver-staining for signal enhancement. Conclusion: A stepwise silver-staining strategy could be useful in minimizing nonspecific background signals. This 96-well-plate-based A beta antibody detection assay could be useful in studying and diagnosing Alzheimer's disease.

引用

页码：485 / 493

页数：9

共 16 条

[1] Colorimetric silver detection of DNA microarrays [J].