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Int. J. Mol. Sci. 2017, 18(2), 470; doi:10.3390/ijms18020470

An Automated Micro-Total Immunoassay System for Measuring Cancer-Associated α2,3-linked Sialyl N-Glycan-Carrying Prostate-Specific Antigen May Improve the Accuracy of Prostate Cancer Diagnosis

1
Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan
2
Diagnostics Research Laboratories, Wako Pure Chemical Industries, Hyogo 661-0963, Japan
3
Department of Advanced Transplant and Regenerative Medicine, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan
4
Department of Urology, Akita University Graduate School of Medicine, Akita 010-8543, Japan
5
Department of Urology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan
6
Department of Surgery, McMaster University, Hamilton, ON L8S4L8, Canada
*
Author to whom correspondence should be addressed.
Academic Editor: Carsten Stephan
Received: 1 February 2017 / Revised: 17 February 2017 / Accepted: 18 February 2017 / Published: 22 February 2017
(This article belongs to the Special Issue Diagnostic, Prognostic and Predictive Biomarkers in Prostate Cancer)
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Abstract

The low specificity of the prostate-specific antigen (PSA) for early detection of prostate cancer (PCa) is a major issue worldwide. The aim of this study to examine whether the serum PCa-associated α2,3-linked sialyl N-glycan-carrying PSA (S2,3PSA) ratio measured by automated micro-total immunoassay systems (μTAS system) can be applied as a diagnostic marker of PCa. The μTAS system can utilize affinity-based separation involving noncovalent interaction between the immunocomplex of S2,3PSA and Maackia amurensis lectin to simultaneously determine concentrations of free PSA and S2,3PSA. To validate quantitative performance, both recombinant S2,3PSA and benign-associated α2,6-linked sialyl N-glycan-carrying PSA (S2,6PSA) purified from culture supernatant of PSA cDNA transiently-transfected Chinese hamster ovary (CHO)-K1 cells were used as standard protein. Between 2007 and 2016, fifty patients with biopsy-proven PCa were pair-matched for age and PSA levels, with the same number of benign prostatic hyperplasia (BPH) patients used to validate the diagnostic performance of serum S2,3PSA ratio. A recombinant S2,3PSA- and S2,6PSA-spiked sample was clearly discriminated by μTAS system. Limit of detection of S2,3PSA was 0.05 ng/mL and coefficient variation was less than 3.1%. The area under the curve (AUC) for detection of PCa for the S2,3PSA ratio (%S2,3PSA) with cutoff value 43.85% (AUC; 0.8340) was much superior to total PSA (AUC; 0.5062) using validation sample set. Although the present results are preliminary, the newly developed μTAS platform for measuring %S2,3PSA can achieve the required assay performance specifications for use in the practical and clinical setting and may improve the accuracy of PCa diagnosis. Additional validation studies are warranted. View Full-Text
Keywords: prostate-specific antigen; α2,3-linked sialyl N-glycan; Maackia amurensis lectin (MAA) lectin; biomarker prostate-specific antigen; α2,3-linked sialyl N-glycan; Maackia amurensis lectin (MAA) lectin; biomarker
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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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Ishikawa, T.; Yoneyama, T.; Tobisawa, Y.; Hatakeyama, S.; Kurosawa, T.; Nakamura, K.; Narita, S.; Mitsuzuka, K.; Duivenvoorden, W.; Pinthus, J.H.; Hashimoto, Y.; Koie, T.; Habuchi, T.; Arai, Y.; Ohyama, C. An Automated Micro-Total Immunoassay System for Measuring Cancer-Associated α2,3-linked Sialyl N-Glycan-Carrying Prostate-Specific Antigen May Improve the Accuracy of Prostate Cancer Diagnosis. Int. J. Mol. Sci. 2017, 18, 470.

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