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

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• 作者：Wanjuan Lin ; Yi Hou ; Yijie Lu ; Ahmed I. Abdelrahman ; Pengpeng Cao ; Guangyao Zhao ; Lemuel Tong ; Jieshu Qian ; Vladimir Baranov ; Mark Nitz ; Mitchell A. Winnik
• 刊名：Langmuir
• 出版年：2014
• 出版时间：March 25, 2014
• 年：2014
• 卷：30
• 期：11
• 页码：3142-3153
• 全文大小：669K
• 年卷期：v.30,no.11(March 25, 2014)
• ISSN：1520-5827

文摘

This paper addresses the question of whether one can use lanthanide nanoparticles (e.g., NaHoF₄) 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 鈥渃ell鈥?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 (>10⁴) 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 Tb³⁺ ions per probe. The other was a biotinylated NaHoF₄ NP containing 15鈥?00 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鈥?00 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.