title: 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.
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 sialylN-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 andMaackia amurensislectin to simultaneously determine concentrations of free PSA and S2,3PSA. To validate quantitative performance, both recombinant S2,3PSA and benign-associated α2,6-linked sialylN-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.
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. ishikawa.tomokazu@wako-chem.co.jp.|Diagnostics Research Laboratories, Wako Pure Chemical Industries, Hyogo 661-0963, Japan. ishikawa.tomokazu@wako-chem.co.jp.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Ishikawa
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. tohruyon@hirosaki-u.ac.jp.|Department of Advanced Transplant and Regenerative Medicine, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. tohruyon@hirosaki-u.ac.jp.
authors.forename: Tohru
authors.initials: T
authors.lastname: Yoneyama
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. tobisawa@hirosaki-u.ac.jp.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tobisawa
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. shingoh@hirosaki-u.ac.jp.
authors.forename: Shingo
authors.initials: S
authors.lastname: Hatakeyama
authors.affiliation: Diagnostics Research Laboratories, Wako Pure Chemical Industries, Hyogo 661-0963, Japan. kurosawa.tatsuo@wako-chem.co.jp.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kurosawa
authors.affiliation: Diagnostics Research Laboratories, Wako Pure Chemical Industries, Hyogo 661-0963, Japan. nakamura.kenji@wako-chem.co.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: Department of Urology, Akita University Graduate School of Medicine, Akita 010-8543, Japan. narishin@doc.med.akita-u.ac.jp.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Narita
authors.affiliation: Department of Urology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan. mitsuzuka@uro.med.tohoku.ac.jp.
authors.forename: Koji
authors.initials: K
authors.lastname: Mitsuzuka
authors.affiliation: Department of Surgery, McMaster University, Hamilton, ON L8S4L8, Canada. duiven@mcmaster.ca.
authors.forename: Wilhelmina
authors.initials: W
authors.lastname: Duivenvoorden
authors.affiliation: Department of Surgery, McMaster University, Hamilton, ON L8S4L8, Canada. pinthusj@hhsc.ca.
authors.forename: Jehonathan H
authors.initials: JH
authors.lastname: Pinthus
authors.affiliation: Department of Advanced Transplant and Regenerative Medicine, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. bikkuri@opal.plala.or.jp.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Hashimoto
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. goodwin@hirosaki-u.ac.jp.
authors.forename: Takuya
authors.initials: T
authors.lastname: Koie
authors.affiliation: Department of Urology, Akita University Graduate School of Medicine, Akita 010-8543, Japan. thabuchi@doc.med.akita-u.ac.jp.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Habuchi
authors.affiliation: Department of Urology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan. yarai@uro.med.tohoku.ac.jp.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. coyama@hirosaki-u.ac.jp.|Department of Advanced Transplant and Regenerative Medicine, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan. coyama@hirosaki-u.ac.jp.
authors.forename: Chikara
authors.initials: C
authors.lastname: Ohyama
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D014408:Biomarkers, Tumor; D016466:CHO Cells; D003412:Cricetulus; D055088:Early Detection of Cancer / Q000379:methods / Q000592:standards; D006031:Glycosylation; D006801:Humans; D007118:Immunoassay; D037102:Lectins; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000379:methods / Q000592:standards; D008875:Middle Aged; D011134:Polysaccharides / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000097:blood* / Q000737:chemistry*; D011471:Prostatic Neoplasms / Q000097:blood* / Q000175:diagnosis*; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D037102:Lectins; D011134:Polysaccharides; D011994:Recombinant Proteins; D017430:Prostate-Specific Antigen
keywords: Maackia amurensis lectin (MAA) lectin; biomarker; prostate-specific antigen; α2,3-linked sialyl N-glycan