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Bioengineering 2019, 6(1), 23; https://doi.org/10.3390/bioengineering6010023

Lanthanide-Loaded Nanoparticles as Potential Fluorescent and Mass Probes for High-Content Protein Analysis

1
Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR 97239, USA
2
PDX Pharmaceuticals, LLC, Portland, OR 97239, USA
3
Department of Biochemistry, Mahidol University, Bangkok 10700, Thailand
*
Author to whom correspondence should be addressed.
Received: 11 February 2019 / Revised: 4 March 2019 / Accepted: 7 March 2019 / Published: 15 March 2019
(This article belongs to the Collection Nanoparticles in Therapeutic Applications)
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Abstract

Multiparametric and high-content protein analysis of single cells or tissues cannot be accomplished with the currently available flow cytometry or imaging techniques utilizing fluorophore-labelled antibodies, because the number of spectrally resolvable fluorochromes is limited. In contrast, mass cytometry can resolve more signals by exploiting lanthanide-tagged antibodies; however, only about 100 metal reporters can be attached to an antibody molecule. This makes the sensitivity of lanthanide-tagged antibodies substantially lower than fluorescent reporters. A new probe that can carry more lanthanide molecules per antibody is a desirable way to enhance the sensitivity needed for the detection of protein with low cellular abundance. Herein, we report on the development of new probes utilizing mesoporous silica nanoparticles (MSNPs) with hydroxyl, amine, or phosphonate functional groups. The phosphonated MSNPs proved to be best at loading lanthanides for up to 1.4 × 106 molecules per particle, and could be loaded with various lanthanide elements (Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, and Lu) at relatively similar molar extents. The modified MSNPs can also load a fluorescent dye, allowing bimodal mass and fluorescence-based detection. We achieved specificity of antibody-conjugated nanoparticles (at 1.4 × 103 antibodies per nanoparticle) for targeting proteins on the cell surface. The new materials can potentially be used as mass cytometry probes and provide a method for simultaneous monitoring of a large host of factors comprising the tumor microenvironment (e.g., extracellular matrix, cancer cells, and immune cells). These novel probes may also benefit personalized medicine by allowing for high-throughput analysis of multiple proteins in the same specimen. View Full-Text
Keywords: lanthanide; nanoparticle; imaging probe; mass cytometry; protein analysis lanthanide; nanoparticle; imaging probe; mass cytometry; protein analysis
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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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Ngamcherdtrakul, W.; Sangvanich, T.; Goodyear, S.; Reda, M.; Gu, S.; Castro, D.J.; Punnakitikashem, P.; Yantasee, W. Lanthanide-Loaded Nanoparticles as Potential Fluorescent and Mass Probes for High-Content Protein Analysis. Bioengineering 2019, 6, 23.

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