A Highly Sensitive Porous Silicon (P-Si)-Based Human Kallikrein 2 (hK2) Immunoassay Platform toward Accurate Diagnosis of Prostate Cancer
| Autor: | Kazuo Hosokawa, Hans Lilja, Mizuo Maeda, Thomas Laurell, Ok Chan Jeong, Sang W ook Lee, Soyoun Kim |
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| Přispěvatelé: | BioMediTech - BioMediTech, University of Tampere |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Male
Silicon Biolääketieteet - Biomedicine chemistry.chemical_element lcsh:Chemical technology Porous silicon Biochemistry Article Analytical Chemistry human kallikrein 2 Adsorption Syöpätaudit - Cancers medicine antibody microarrays Biomarkers Tumor Humans lcsh:TP1-1185 Electrical and Electronic Engineering Instrumentation Detection limit Immunoassay Chromatography biology medicine.diagnostic_test Chemistry Nanoporous Prostatic Neoplasms Equipment Design prostate cancer Microarray Analysis Molecular biology Atomic and Molecular Physics and Optics porous silicon biology.protein Titration Antibody Medicinal Chemistry Porosity Tissue Kallikreins |
| Zdroj: | Sensors (Basel, Switzerland) Sensors; 15(5), pp 11972-11987 (2015) Sensors, Vol 15, Iss 5, Pp 11972-11987 (2015) Sensors Volume 15 Issue 5 Pages 11972-11987 |
| ISSN: | 1424-8220 |
| Popis: | 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 10(2) ng/mL). |
| Databáze: | OpenAIRE |
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