Cargando…

A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells

[Image: see text] This paper addresses the question of whether one can use lanthanide nanoparticles (e.g., NaHoF(4)) to detect surface biomarkers expressed at low levels by mass cytometry. To avoid many of the complications of experiments on live or fixed cells, we carried out proof-of-concept exper...

Descripción completa

Detalles Bibliográficos
Autores principales: Lin, Wanjuan, Hou, Yi, Lu, Yijie, Abdelrahman, Ahmed I., Cao, Pengpeng, Zhao, Guangyao, Tong, Lemuel, Qian, Jieshu, Baranov, Vladimir, Nitz, Mark, Winnik, Mitchell A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3983384/
https://www.ncbi.nlm.nih.gov/pubmed/24617504
http://dx.doi.org/10.1021/la403627p
_version_ 1782311318248751104
author Lin, Wanjuan
Hou, Yi
Lu, Yijie
Abdelrahman, Ahmed I.
Cao, Pengpeng
Zhao, Guangyao
Tong, Lemuel
Qian, Jieshu
Baranov, Vladimir
Nitz, Mark
Winnik, Mitchell A.
author_facet Lin, Wanjuan
Hou, Yi
Lu, Yijie
Abdelrahman, Ahmed I.
Cao, Pengpeng
Zhao, Guangyao
Tong, Lemuel
Qian, Jieshu
Baranov, Vladimir
Nitz, Mark
Winnik, Mitchell A.
author_sort Lin, Wanjuan
collection PubMed
description [Image: see text] This paper addresses the question of whether one can use lanthanide nanoparticles (e.g., NaHoF(4)) to detect surface biomarkers expressed at low levels by mass cytometry. To avoid many of the complications of experiments on live or fixed cells, we carried out proof-of-concept experiments using aqueous microgels with a diameter on the order of 700 nm as a proxy for cells. These microgels were used to test whether nanoparticle (NP) reagents would allow the detection of as few as 100 proteins per “cell” in cell-by-cell assays. Streptavidin (SAv), which served as the model biomarker, was attached to the microgel in two different ways. Covalent coupling to surface carboxyls of the microgel led to large numbers (>10(4)) of proteins per microgel, whereas biotinylation of the microgel followed by exposure to SAv led to much smaller numbers of SAv per microgel. Using mass cytometry, we compared two biotin-containing reagents, which recognized and bound to the SAvs on the microgel. One was a metal chelating polymer (MCP), a biotin end-capped polyaspartamide containing 50 Tb(3+) ions per probe. The other was a biotinylated NaHoF(4) NP containing 15 000 Ho atoms per probe. Nonspecific binding was determined with bovine serum albumin (BSA) conjugated microgels. The MCP was effective at detecting and quantifying SAvs on the microgel with covalently bound SAv (20 000 SAvs per microgel) but was unable to give a meaningful signal above that of the BSA-coated microgel for the samples with low levels of SAv. Here the NP reagent gave a signal 2 orders of magnitude stronger than that of the MCP and allowed detection of NPs ranging from 100 to 500 per microgel. Sensitivity was limited by the level of nonspecific adsorption. This proof of concept experiment demonstrates the enhanced sensitivity possible with NP reagents in cell-by-cell assays by mass cytometry.
format Online
Article
Text
id pubmed-3983384
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-39833842015-03-11 A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells Lin, Wanjuan Hou, Yi Lu, Yijie Abdelrahman, Ahmed I. Cao, Pengpeng Zhao, Guangyao Tong, Lemuel Qian, Jieshu Baranov, Vladimir Nitz, Mark Winnik, Mitchell A. Langmuir [Image: see text] This paper addresses the question of whether one can use lanthanide nanoparticles (e.g., NaHoF(4)) to detect surface biomarkers expressed at low levels by mass cytometry. To avoid many of the complications of experiments on live or fixed cells, we carried out proof-of-concept experiments using aqueous microgels with a diameter on the order of 700 nm as a proxy for cells. These microgels were used to test whether nanoparticle (NP) reagents would allow the detection of as few as 100 proteins per “cell” in cell-by-cell assays. Streptavidin (SAv), which served as the model biomarker, was attached to the microgel in two different ways. Covalent coupling to surface carboxyls of the microgel led to large numbers (>10(4)) of proteins per microgel, whereas biotinylation of the microgel followed by exposure to SAv led to much smaller numbers of SAv per microgel. Using mass cytometry, we compared two biotin-containing reagents, which recognized and bound to the SAvs on the microgel. One was a metal chelating polymer (MCP), a biotin end-capped polyaspartamide containing 50 Tb(3+) ions per probe. The other was a biotinylated NaHoF(4) NP containing 15 000 Ho atoms per probe. Nonspecific binding was determined with bovine serum albumin (BSA) conjugated microgels. The MCP was effective at detecting and quantifying SAvs on the microgel with covalently bound SAv (20 000 SAvs per microgel) but was unable to give a meaningful signal above that of the BSA-coated microgel for the samples with low levels of SAv. Here the NP reagent gave a signal 2 orders of magnitude stronger than that of the MCP and allowed detection of NPs ranging from 100 to 500 per microgel. Sensitivity was limited by the level of nonspecific adsorption. This proof of concept experiment demonstrates the enhanced sensitivity possible with NP reagents in cell-by-cell assays by mass cytometry. American Chemical Society 2014-03-11 2014-03-25 /pmc/articles/PMC3983384/ /pubmed/24617504 http://dx.doi.org/10.1021/la403627p Text en Copyright © 2014 American Chemical Society
spellingShingle Lin, Wanjuan
Hou, Yi
Lu, Yijie
Abdelrahman, Ahmed I.
Cao, Pengpeng
Zhao, Guangyao
Tong, Lemuel
Qian, Jieshu
Baranov, Vladimir
Nitz, Mark
Winnik, Mitchell A.
A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title_full A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title_fullStr A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title_full_unstemmed A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title_short A High-Sensitivity Lanthanide Nanoparticle Reporter for Mass Cytometry: Tests on Microgels as a Proxy for Cells
title_sort high-sensitivity lanthanide nanoparticle reporter for mass cytometry: tests on microgels as a proxy for cells
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3983384/
https://www.ncbi.nlm.nih.gov/pubmed/24617504
http://dx.doi.org/10.1021/la403627p
work_keys_str_mv AT linwanjuan ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT houyi ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT luyijie ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT abdelrahmanahmedi ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT caopengpeng ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT zhaoguangyao ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT tonglemuel ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT qianjieshu ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT baranovvladimir ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT nitzmark ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT winnikmitchella ahighsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT linwanjuan highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT houyi highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT luyijie highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT abdelrahmanahmedi highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT caopengpeng highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT zhaoguangyao highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT tonglemuel highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT qianjieshu highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT baranovvladimir highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT nitzmark highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells
AT winnikmitchella highsensitivitylanthanidenanoparticlereporterformasscytometrytestsonmicrogelsasaproxyforcells