Platinum-conjugated Antibodies for Application in Mass Cytometry
Platinum-conjugated Antibodies for Application in Mass Cytometry
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DOI：
10.1002/cyto.a.22778
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发表时间：
2016-03-01
期刊：
影响因子：
3.7
通讯作者：
Maecker, Holden T.
中科院分区：
文献类型：
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作者：
Mei, Henrik E.;Leipold, Michael D.;Maecker, Holden T.
质谱细胞术已经克服了荧光单细胞细胞术的局限性，允许在单细胞水平上测量多达40个不同的参数。然而，细胞蛋白质组包含更多潜在的分析物，并且当前的质谱细胞术仪器理论上允许多达121个不同的质量检测通道。用适当的金属离子标记特异性探针是利用更多的质谱细胞术的分析能力的一个重要障碍。为此，我们在这里描述了标记抗体与天然丰度或同位素纯化的铂配制在顺铂和规避使用螯合剂负载的聚合物。我们确认了顺铂-抗体-缀合物用于表面、细胞内和磷酸化表位特异性免疫表型分析的效用，以及用于基于细胞表面CD 45的条形码化的应用。顺铂标记抗体使CyTOF（R）平台的分析能力基于现有试剂增加了两个通道，并有可能增加总共六个通道用于检测特异性探针，从而有助于更好地扩展质谱流式细胞仪的分析质量范围。(C)2015年国际细胞计数促进学会
Mass cytometry has overcome limitations of fluorescent single cell cytometry by allowing for the measurement of up to currently similar to 40 different parameters on a single cell level. However, the cellular proteome comprises many more potential analytes, and current mass cytometry instrumentation allows for theoretically up to 121 different mass detection channels. The labeling of specific probes with appropriate metal ions is a significant hurdle for exploiting more of mass cytometry's analytical capacity. To this end, we here describe the labeling of antibody with natural abundance or isotopically purified platinum as formulated in cisplatin and circumventing the use of chelator-loaded polymers. We confirm the utility of cisplatin-antibody-conjugates for surface, intracellular, and phosphoepitope-specific immunophenotyping, as well as for application in cell surface CD45-based barcoding. Cisplatin-labeling of antibody increases the analytical capacity of the CyTOF (R) platform by two channels based on available reagents, and has the potential to add a total of six channels for detection of specific probes, thus helping to better extend the analytical mass range of mass cytometers. (C) 2015 International Society for Advancement of Cytometry