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Improved porous silicon microarray based prostate specific antigen immunoassay by optimized surface density of the capture antibody
Authors:SangWook Lee  Soyoun Kim  Johan Malm  Ok Chan Jeong  Hans Lilja  Thomas Laurell
Institution:1. Bioengineering Laboratory, Riken Institute, Saitama, Japan;2. Department of Biomedical Engineering, Dongguk University, Seoul, Republic of Korea;3. Department of Laboratory Medicine, Division of Clinical Chemistry, Lund University, Skåne University Hospital in Malmö, 205 02 Malmö, Sweden;4. Department of Mechanical Engineering, Inje University, Busan, Republic of Korea;5. Department of Laboratory Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY, United States;6. Department of Surgery (Urology), Memorial Sloan-Kettering Cancer Center, New York, NY, United States;g Department of Medicine (GU Oncology), Memorial Sloan-Kettering Cancer Center, New York, NY, United States;h Institute for Biosciences and Medical Technology, University of Tampere, Tampere, Finland;i Nuffield Department of Surgical Sciences, University of Oxford, Oxford, United Kingdom;j Department of Electrical Measurements, Lund University, Box 118, 221 00 Lund, Sweden
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.
Keywords:Antibody microarray  Porous silicon  Sandwich immunoassay  Prostate specific antigen
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