Takahashi Chikara, Au-Yeung Amelia, Fuh Franklin, Ramirez-Montagut Teresa, Bolen Chris, Mathews William, O'Gorman William E
OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
Bioinformatics Department, Genentech, South San Francisco, California, 94080.
Cytometry A. 2017 Jan;91(1):39-47. doi: 10.1002/cyto.a.22977. Epub 2016 Sep 15.
Mass cytometry is capable of measuring more than 40 distinct proteins on individual cells making it a promising technology for innovating biomarker discovery. However, in order for this potential to be fully realized, best practices in panel design need to be further defined in order to achieve consistency and reproducibility in data analysis. Of particular importance are controls that reveal, and panel design principles that mitigate the effects of signal interference or overlap. We observed a disparity between the staining profiles of two noncompeting anti- integrin β7 mAbs and hypothesized that signal interference was responsible. A mass-minus-one (MMO) control was applied and demonstrated that signal overlap caused the perceived interclonal discrepancy in β7 expression. Panel redesign in consideration of mass-cytometry specific interference dynamics dramatically improved concordance between both mAbs by redistributing background signals caused by overlap. These studies visualize how signal overlap can complicate mass cytometry data interpretation and demonstrate how the rational distribution of interference can greatly improve panel design and data quality. © 2016 International Society for Advancement of Cytometry.
质谱流式细胞术能够对单个细胞上的40多种不同蛋白质进行测量，使其成为创新生物标志物发现的一项有前景的技术。然而，为了充分实现这一潜力，需要进一步明确分组设计的最佳实践，以便在数据分析中实现一致性和可重复性。特别重要的是能够揭示问题的对照以及能够减轻信号干扰或重叠影响的分组设计原则。我们观察到两种非竞争性抗整合素β7单克隆抗体的染色谱存在差异，并推测是信号干扰所致。应用了质量减一（MMO）对照，结果表明信号重叠导致了β7表达中观察到的克隆间差异。考虑到质谱流式细胞术特有的干扰动态进行分组重新设计，通过重新分配由重叠引起的背景信号，显著提高了两种单克隆抗体之间的一致性。这些研究直观展示了信号重叠如何使质谱流式细胞术数据解读变得复杂，并证明了干扰的合理分布如何能够极大地改进分组设计和数据质量。© 2016国际细胞计量学促进协会。
