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Sensors 2015, 15(5), 11972-11987; doi:10.3390/s150511972

A Highly Sensitive Porous Silicon (P-Si)-Based Human Kallikrein 2 (hK2) Immunoassay Platform toward Accurate Diagnosis of Prostate Cancer

1
Bioengineering Laboratory, RIKEN, Saitama 3510198, Japan
2
Department of Biomedical Engineering, Dongguk University, Seoul 100715, Korea
3
Department of Mechanical Engineering, Inje University, Gimhae-si 621-749, Korea
4
Department of Translational Medicine, Lund University, Skåne University Hospital in Malmö, Malmö 20502, Sweden
5
Departments of Laboratory Medicine, Surgery (Urology), and Medicine (GU Oncology), Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA
6
Institute for Biosciences and Medical Technology, University of Tampere, Tampere 33800, Finland
7
Nuffield Department of Surgical Sciences, University of Oxford, Oxford OX3 7DQ, UK
8
Department Biomedical Engineering, Lund University, Lund 22100, Sweden
*
Authors to whom correspondence should be addressed.
Academic Editor: Alexander Star
Received: 19 February 2015 / Accepted: 14 May 2015 / Published: 22 May 2015
(This article belongs to the Special Issue Immunosensors 2014)
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Abstract

Levels of total human kallikrein 2 (hK2), a protein involved the pathology of prostate cancer (PCa), could be used as a biomarker to aid in the diagnosis of this disease. In this study, we report on a porous silicon antibody immunoassay platform for the detection of serum levels of total hK2. The surface of porous silicon has a 3-dimensional macro- and nanoporous structure, which offers a large binding capacity for capturing probe molecules. The tailored pore size of the porous silicon also allows efficient immobilization of antibodies by surface adsorption, and does not require chemical immobilization. Monoclonal hK2 capture antibody (6B7) was dispensed onto P-Si chip using a piezoelectric dispenser. In total 13 × 13 arrays (169 spots) were spotted on the chip with its single spot volume of 300 pL. For an optimization of capture antibody condition, we firstly performed an immunoassay of the P-Si microarray under a titration series of hK2 in pure buffer (PBS) at three different antibody densities (75, 100 and 145 µg/mL). The best performance of the microarray platform was seen at 100 µg/mL of the capture antibody concentration (LOD was 100 fg/mL). The platform then was subsequently evaluated for a titration series of serum-spiked hK2 samples. The developed platform utilizes only 15 µL of serum per test and the total assay time is about 3 h, including immobilization of the capture antibody. The detection limit of the hK2 assay was 100 fg/mL in PBS buffer and 1 pg/mL in serum with a dynamic range of 106 (10−4 to 102 ng/mL). View Full-Text
Keywords: porous silicon; human kallikrein 2; antibody microarrays; prostate cancer porous silicon; human kallikrein 2; antibody microarrays; prostate cancer
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Lee, S.W.; Hosokawa, K.; Kim, S.; Jeong, O.C.; Lilja, H.; Laurell, T.; Maeda, M. A Highly Sensitive Porous Silicon (P-Si)-Based Human Kallikrein 2 (hK2) Immunoassay Platform toward Accurate Diagnosis of Prostate Cancer. Sensors 2015, 15, 11972-11987.

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