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
:1. Introduction
2. Results
2.1. Preparation of FLAG-Tag-Fused Recombinat S2,3 and S2,6 PSA in Chinese Hamster Ovary (CHO) Cells
2.2. S2,6PSA and S2,3PSA Separation in a Microfluidic Channel Filled with Electrophoresis Leading Buffer Containing Maackia Amurensis Lectin
2.3. Assay Linearity and Sensitivity of %S2,3PSA Test by μTAS System
2.4. Assay Reproducibility of %S2,3PSA Test Using the μTAS System
2.5. Validation of %S2,3PSA Test
3. Discussion
4. Materials and Methods
4.1. Immunoassay Reagents
4.2. Microfluidic Electrophoresis Assay
4.3. Prostate Biopsy and Serum Samples
4.4. Forced Expression of FLAG-Tag-Fused S2,3 and S2,6PSA in Chinese Hamster Ovary (CHO)-K1 Cells
4.5. Lectin Microarray
4.6. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
μTAS | micro-total analysis system |
MAA | maackia amurensis lectin |
EATA | electrokinetic analyte transport assay |
LBA | liquid-phase binding assay |
C.V. | coefficient of variation |
ITP | isotachophoresis |
LIF | laser-induced fluorescence |
LB | leading buffer |
TB | trailing buffer |
CE | capillary electrophoresis |
SB | sample buffer |
ST | stacking buffer |
CGE | capillary gel electrophoresis |
LOD | limit of detection |
PSA | prostate-specific antigen |
PCa | prostate cancer |
BPH | benign prostatic hyperplasia |
Pbx GG | prostate biopsy grade group |
LacdiNAc | GalNAcβ1-4GlcNAc- |
Gal | galactose |
Man | mannose |
Fuc | fucose |
Sia | sialic acid |
GalNAc | N-acetylgalactosamine |
GlcNAc | N-acetylglucosamine |
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Sample Number | 1.0 ng/mL Free PSA | 5.0 ng/mL Free PSA | ||
---|---|---|---|---|
Free PSA | %S2,3PSA | Free PSA | %S2,3PSA | |
1 | 1.04 | 50.5 | 5.06 | 38.1 |
2 | 0.99 | 50.7 | 5.08 | 38.1 |
3 | 1.04 | 49.4 | 5.00 | 38.3 |
4 | 1.05 | 50.1 | 5.07 | 37.9 |
5 | 1.04 | 47.7 | 5.05 | 37.7 |
6 | 1.03 | 50.4 | 5.24 | 37.7 |
7 | 1.13 | 50.3 | 5.13 | 38.3 |
8 | 1.10 | 46.8 | 5.01 | 37.9 |
9 | 1.05 | 47.1 | 5.04 | 38.1 |
10 | 1.01 | 48.5 | 5.21 | 37.9 |
Ave. 1 | 1.04 | 49.1 | 5.09 | 38.0 |
SD 2 | 0.03 | 1.51 | 0.08 | 0.21 |
CV 3 | 2.8% | 3.1% | 1.6% | 0.6% |
Characteristics | BPH a | PCa b | p (a vs. b) |
---|---|---|---|
n = 100 | 50 | 50 | |
Age, median (range) | 66.5 (51–85) | 67 (51–86) | ns 1 |
PSA 2, ng/mL, median (range) | 6.45 (1.9–20.4) | 6.6 (1.5–21.4) | ns 1 |
%S2,3PSA, median (range) | 38.55 (22.9–59.1) | 45.70 (34.7–71.7) | <0.0001 |
pbx GG 3 | n (%) | ||
GG 1 | 15 (30%) | ||
GG 2 | 12 (24%) | ||
GG 3 | 8 (16%) | ||
GG 4 | 9 (18%) | ||
GG 5 | 6 (12%) |
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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.; et al. 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. https://doi.org/10.3390/ijms18020470
Ishikawa T, Yoneyama T, Tobisawa Y, Hatakeyama S, Kurosawa T, Nakamura K, Narita S, Mitsuzuka K, Duivenvoorden W, Pinthus JH, et al. 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. International Journal of Molecular Sciences. 2017; 18(2):470. https://doi.org/10.3390/ijms18020470
Chicago/Turabian StyleIshikawa, Tomokazu, Tohru Yoneyama, Yuki Tobisawa, Shingo Hatakeyama, Tatsuo Kurosawa, Kenji Nakamura, Shintaro Narita, Koji Mitsuzuka, Wilhelmina Duivenvoorden, Jehonathan H. Pinthus, and et al. 2017. "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" International Journal of Molecular Sciences 18, no. 2: 470. https://doi.org/10.3390/ijms18020470