title: Improved porous silicon microarray based prostate specific antigen immunoassay by optimized surface density of the capture antibody.
abstract: Enriching the surface density of immobilized capture antibodies enhances the detection signal of antibody sandwich microarrays. In this study, we improved the detection sensitivity of our previously developed P-Si (porous silicon) antibody microarray by optimizing concentrations of the capturing antibody. We investigated immunoassays using a P-Si microarray at three different capture antibody (PSA - prostate specific antigen) concentrations, analyzing the influence of the antibody density on the assay detection sensitivity. The LOD (limit of detection) for PSA was 2.5 ng mL(-1), 80 pg mL(-1), and 800 fg mL(-1) when arraying the PSA antibody, H117 at the concentration 15 μg mL(-1), 35 μg mL(-1), and 154 μg mL(-1), respectively. We further investigated PSA spiked into human female serum in the range of 800 fg mL(-1) to 500 ng mL(-1). The microarray showed a LOD of 800 fg mL(-1) and a dynamic range of 800 fg mL(-1) to 80 ng mL(-1) in serum spiked samples.
authors.affiliation: Bioengineering Laboratory, Riken Institute, Saitama, Japan; Department of Biomedical Engineering, Dongguk University, Seoul, Republic of Korea. Electronic address: Sangwook_L@riken.jp.
authors.forename: SangWook
authors.initials: S
authors.lastname: Lee
authors.forename: Soyoun
authors.initials: S
authors.lastname: Kim
authors.forename: Johan
authors.initials: J
authors.lastname: Malm
authors.forename: Ok Chan
authors.initials: OC
authors.lastname: Jeong
authors.forename: Hans
authors.initials: H
authors.lastname: Lilja
authors.forename: Thomas
authors.initials: T
authors.lastname: Laurell
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D004867:Equipment Design; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D057230:Limit of Detection; D008297:Male; D016062:Porosity; D017430:Prostate-Specific Antigen / Q000097:blood*; D040081:Protein Array Analysis / Q000295:instrumentation*; D012825:Silicon / Q000737:chemistry*
chemical_list: D055910:Antibodies, Immobilized; D017430:Prostate-Specific Antigen; D012825:Silicon
keywords: Antibody microarray; Porous silicon; Prostate specific antigen; Sandwich immunoassay
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