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Nanomedicine (Lond). 2008 Aug;3(4):485-93. doi: 10.2217/17435889.3.4.485.

Stepwise silver-staining-based immunosorbent assay for amyloid-beta autoantibody detection.

Author information

1
Department of Chemistry, Seoul National University, Seoul, South Korea.

Abstract

AIMS:

To detect amyloid-beta (Abeta) 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:

Abeta autoantibodies were incubated in an Abeta-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 Abeta antibody detection assay could be useful in studying and diagnosing Alzheimer's disease.

PMID:
18694311
DOI:
10.2217/17435889.3.4.485
[Indexed for MEDLINE]

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