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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.

Original publication

DOI

10.1016/j.aca.2013.06.041

Type

Journal article

Journal

Anal Chim Acta

Publication Date

24/09/2013

Volume

796

Pages

108 - 114

Keywords

Antibody microarray, Porous silicon, Prostate specific antigen, Sandwich immunoassay, Antibodies, Immobilized, Equipment Design, Female, Humans, Immunoassay, Limit of Detection, Male, Porosity, Prostate-Specific Antigen, Protein Array Analysis, Silicon