Improved porous silicon microarray based prostate specific antigen immunoassay by optimized surface density of the capture antibody

被引:19
作者
Lee, SangWook [1 ,2 ]
Kim, Soyoun [2 ]
Malm, Johan [3 ]
Jeong, Ok Chan [4 ]
Lilja, Hans [3 ,5 ,6 ,7 ,8 ,9 ]
Laurell, Thomas [2 ,10 ]
机构
[1] Riken Inst Phys & Chem Res, Bioengn Lab, Saitama, Japan
[2] Dongguk Univ, Dept Biomed Engn, Seoul, South Korea
[3] Lund Univ, Skane Univ Hosp Malmo, Div Clin Chem, Dept Lab Med, S-20502 Malmo, Sweden
[4] Inje Univ, Dept Mech Engn, Pusan, South Korea
[5] Mem Sloan Kettering Canc Ctr, Dept Lab Med, New York, NY 10021 USA
[6] Mem Sloan Kettering Canc Ctr, Dept Surg Urol, New York, NY 10021 USA
[7] Mem Sloan Kettering Canc Ctr, Dept Med GU Oncol, New York, NY 10021 USA
[8] Univ Tampere, Inst Biosci & Med Technol, FIN-33101 Tampere, Finland
[9] Univ Oxford, Nuffield Dept Surg Sci, Oxford, England
[10] Lund Univ, Dept Elect Measurements, S-22100 Lund, Sweden
基金
瑞典研究理事会;
关键词
Antibody microarray; Porous silicon; Sandwich immunoassay; Prostate specific antigen; PHASE PROTEIN MICROARRAYS; REVERSE-PHASE; BIOCHEMICAL RECURRENCE; RADICAL PROSTATECTOMY; CANCER; TECHNOLOGY; SERUM; PSA; AMPLIFICATION; ENHANCEMENT;
D O I
10.1016/j.aca.2013.06.041
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 mu g mL(-1), 35 mu g mL(-1), and 154 mu g 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. (C) 2013 Elsevier B.V. All rights reserved.
引用
收藏
页码:108 / 114
页数:7
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