Upconversion nanoparticle-assisted single-molecule assay for detecting circulating antigens of aggressive prostate cancer.

Chen, Y; Shimoni, O; Huang, G; Wen, S; Liao, J; Duong, HTT; Maddahfar, M; Su, QP; Ortega, DG; Lu, Y; Campbell, DH; Walsh, BJ; Jin, D

Upconversion nanoparticle-assisted single-molecule assay for detecting circulating antigens of aggressive prostate cancer.

Chen, Y

Shimoni, O

Huang, G

Wen, S

Liao, J

Duong, HTT Maddahfar, M Su, QP Ortega, DG Lu, Y Campbell, DH Walsh, BJ Jin, D

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Publisher:: WILEY
Publication Type:: Journal Article
Citation:: Cytometry A, 2022, 101, (5), pp. 400-410
Issue Date:: 2022-01-01

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Field	Value	Language
dc.contributor.author	Chen, Y https://orcid.org/0000-0002-7886-7144
dc.contributor.author	Shimoni, O https://orcid.org/0000-0001-8822-1024
dc.contributor.author	Huang, G https://orcid.org/0000-0001-7524-8199
dc.contributor.author	Wen, S https://orcid.org/0000-0002-4670-4658
dc.contributor.author	Liao, J https://orcid.org/0000-0003-0616-4762
dc.contributor.author	Duong, HTT
dc.contributor.author	Maddahfar, M
dc.contributor.author	Su, QP
dc.contributor.author	Ortega, DG
dc.contributor.author	Lu, Y
dc.contributor.author	Campbell, DH
dc.contributor.author	Walsh, BJ
dc.contributor.author	Jin, D https://orcid.org/0000-0003-1046-2666
dc.date.accessioned	2023-01-25T03:47:55Z
dc.date.available	2021-09-20
dc.date.available	2023-01-25T03:47:55Z
dc.date.issued	2022-01-01
dc.identifier.citation	Cytometry A, 2022, 101, (5), pp. 400-410
dc.identifier.issn	1552-4922
dc.identifier.issn	1552-4930
dc.identifier.uri	http://hdl.handle.net/10453/165430
dc.description.abstract	Sensitive and quantitative detection of molecular biomarkers is crucial for the early diagnosis of diseases like metabolic syndrome and cancer. Here we present a single-molecule sandwich immunoassay by imaging the number of single nanoparticles to diagnose aggressive prostate cancer. Our assay employed the photo-stable upconversion nanoparticles (UCNPs) as labels to detect the four types of circulating antigens in blood circulation, including glypican-1 (GPC-1), leptin, osteopontin (OPN), and vascular endothelial growth factor (VEGF), as their serum concentrations indicate aggressive prostate cancer. Under a wide-field microscope, a single UCNP doped with thousands of lanthanide ions can emit sufficiently bright anti-Stokes' luminescence to become quantitatively detectable. By counting every single streptavidin-functionalized UCNP which specifically labeled on each sandwich immune complex across multiple fields of views, we achieved the Limit of Detection (LOD) of 0.0123 ng/ml, 0.2711 ng/ml, 0.1238 ng/ml, and 0.0158 ng/ml for GPC-1, leptin, OPN and VEGF, respectively. The serum circulating level of GPC-1, leptin, OPN, and VEGF in a mixture of 10 healthy normal human serum was 25.17 ng/ml, 18.04 ng/ml, 11.34 ng/ml, and 1.55 ng/ml, which was within the assay dynamic detection range for each analyte. Moreover, a 20% increase of GPC-1 and OPN was observed by spiking the normal human serum with recombinant antigens to confirm the accuracy of the assay. We observed no cross-reactivity among the four biomarker analytes, which eliminates the false positives and enhances the detection accuracy. The developed single upconversion nanoparticle-assisted single-molecule assay suggests its potential in clinical usage for prostate cancer detection by monitoring tiny concentration differences in a panel of serum biomarkers.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	WILEY
dc.relation	Department of Innovation, Industry, Science and Research (Previously known as DEST)VNSDSHWJ
dc.relation	National Heart Foundation of Australia102592
dc.relation	http://purl.org/au-research/grants/nhmrc/APP1177374
dc.relation	http://purl.org/au-research/grants/arc/IH150100028
dc.relation	http://purl.org/au-research/grants/nhmrc/1101258
dc.relation	http://purl.org/au-research/grants/nhmrc/GNT1177374
dc.relation.ispartof	Cytometry A
dc.relation.isbasedon	10.1002/cyto.a.24504
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0601 Biochemistry and Cell Biology
dc.subject.classification	Immunology
dc.subject.mesh	Biomarkers
dc.subject.mesh	Humans
dc.subject.mesh	Leptin
dc.subject.mesh	Male
dc.subject.mesh	Nanoparticles
dc.subject.mesh	Prostatic Neoplasms
dc.subject.mesh	Vascular Endothelial Growth Factor A
dc.subject.mesh	Humans
dc.subject.mesh	Prostatic Neoplasms
dc.subject.mesh	Leptin
dc.subject.mesh	Vascular Endothelial Growth Factor A
dc.subject.mesh	Male
dc.subject.mesh	Nanoparticles
dc.subject.mesh	Biomarkers
dc.subject.mesh	Biomarkers
dc.subject.mesh	Humans
dc.subject.mesh	Leptin
dc.subject.mesh	Male
dc.subject.mesh	Nanoparticles
dc.subject.mesh	Prostatic Neoplasms
dc.subject.mesh	Vascular Endothelial Growth Factor A
dc.title	Upconversion nanoparticle-assisted single-molecule assay for detecting circulating antigens of aggressive prostate cancer.
dc.type	Journal Article
utslib.citation.volume	101
utslib.location.activity	United States
utslib.for	0601 Biochemistry and Cell Biology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Medical and Molecular Sciences
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
pubs.organisational-group	/University of Technology Sydney/Strength - IBMD - Initiative for Biomedical Devices
pubs.organisational-group	/University of Technology Sydney/Centre for Health Technologies (CHT)
utslib.copyright.status	closed_access	*
pubs.consider-herdc	false
dc.date.updated	2023-01-25T03:47:53Z
pubs.issue	5
pubs.publication-status	Published online
pubs.volume	101
utslib.citation.issue	5

Abstract:

Sensitive and quantitative detection of molecular biomarkers is crucial for the early diagnosis of diseases like metabolic syndrome and cancer. Here we present a single-molecule sandwich immunoassay by imaging the number of single nanoparticles to diagnose aggressive prostate cancer. Our assay employed the photo-stable upconversion nanoparticles (UCNPs) as labels to detect the four types of circulating antigens in blood circulation, including glypican-1 (GPC-1), leptin, osteopontin (OPN), and vascular endothelial growth factor (VEGF), as their serum concentrations indicate aggressive prostate cancer. Under a wide-field microscope, a single UCNP doped with thousands of lanthanide ions can emit sufficiently bright anti-Stokes' luminescence to become quantitatively detectable. By counting every single streptavidin-functionalized UCNP which specifically labeled on each sandwich immune complex across multiple fields of views, we achieved the Limit of Detection (LOD) of 0.0123 ng/ml, 0.2711 ng/ml, 0.1238 ng/ml, and 0.0158 ng/ml for GPC-1, leptin, OPN and VEGF, respectively. The serum circulating level of GPC-1, leptin, OPN, and VEGF in a mixture of 10 healthy normal human serum was 25.17 ng/ml, 18.04 ng/ml, 11.34 ng/ml, and 1.55 ng/ml, which was within the assay dynamic detection range for each analyte. Moreover, a 20% increase of GPC-1 and OPN was observed by spiking the normal human serum with recombinant antigens to confirm the accuracy of the assay. We observed no cross-reactivity among the four biomarker analytes, which eliminates the false positives and enhances the detection accuracy. The developed single upconversion nanoparticle-assisted single-molecule assay suggests its potential in clinical usage for prostate cancer detection by monitoring tiny concentration differences in a panel of serum biomarkers.

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http://hdl.handle.net/10453/165430