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Porous silicon microarray for simultaneous fluorometric immunoassay of the biomarkers prostate-specific antigen and human glandular kallikrein 2

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Abstract

The authors have developed a porous silicon (P-Si) based duplex antibody microarray platform for simultaneous quantitation of the biomarkers prostate-specific antigen (PSA) and human glandular kallikrein 2 (hK2) in serum. Pore size-controlled P-Si surfaces have an extremely enlarged surface area that enables high-density immobilization of fluorescently labeled antibodies by physical adsorption. Automated microarraying of the antibodies provides a fast and reproducible duplex format of antibody arrays on the P-Si chips placed in the wells of a microtiter plate. The assay platform showed a 100 fg·mL−1 limit of detection for both PSA and hK2, and a dynamic range that extends over five orders of magnitude. After optimization of the density of both capture antibodies, neither the PSA nor the hK2 array showed cross-sensitivity to non-target proteins or other plasma proteins. The microarray was evaluated by titration of PSA and hK2, respectively, in the same serum samples. In our perception, this highly sensitive and selective platform holds promise for improved detection of tumor markers in an early diagnostic stage, but also to monitor the recurrence of prostate cancer.

A porous silicon microarray platform is presented for simultaneous detection of biomarkers prostate specific antigen and human glandular kallikrein 2 in human blood sample. The platform shows highly sensitive and selective biomarker detection features.

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Acknowledgments

This study was partly supported by RIKEN FPR program and JSPS KAKENHI Grant Number 25350581. This project is also supported by Korea Ministry of Environment as “EI project” (ERL E211-41003-0007-0).

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Correspondence to SangWook Lee.

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The author(s) declare that they have no competing interests.

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Lee, S., Hosokawa, K., Kim, S. et al. Porous silicon microarray for simultaneous fluorometric immunoassay of the biomarkers prostate-specific antigen and human glandular kallikrein 2. Microchim Acta 183, 3321–3327 (2016). https://doi.org/10.1007/s00604-016-1986-1

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Keywords

  • Duplex antibody microarray
  • PSA
  • hK2
  • Silicon anodization
  • Prostate cancer
  • Serum analysis