Stepwise silver-staining-based immunosorbent assay for amyloid-ββ autoantibody detection. (August 2008)

• Record Type: Journal Article
• Authors: Shim, So-Youn; Woo, Jung-Reem; Nam, Eun-Ju; Hong, Hyun-Joo; Mook-Jung, Inhee; Kim, Young-Ho; Nam, Jwa-Min
• Is Part Of: Nanomedicine. Volume 3:Number 4(2008)
• Page Start: 485
• Page End: 493
• Publication Date: 2008-08
• Subjects: Alzheimer''s disease -- amyloid-ββ autoantibody -- gold nanoparticle -- immunosorbent assay -- silver staining; Nanotechnology -- Periodicals; Medical technology -- Periodicals; Nanotechnology -- Therapeutic use -- Periodicals; 610.28
• DOI: 10.2217/17435889.3.4.485
• Languages: English
• ISSNs: 1743-5889

Abstract

Aims: To detect amyloid-ββ (Aββ) 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: Aββ autoantibodies were incubated in an Aββ-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 Aββ antibody detection assay could be useful in studying and diagnosing Alzheimer''s disease.