normacurve a supercurve-based method that simultaneously quantifies and normalizes reverse phase protein array datanormacurve supercurve-based方法同时量化和规范反向阶段蛋白质阵列数据.pdfVIP

NormaCurve: A SuperCurve-Based Method That Simultaneously Quantifies and Normalizes Reverse Phase Protein Array Data 1,3,4 ´ 2 2 2 2 1 Sylvie Troncale , Aurelie Barbet , Lamine Coulibaly , Emilie Henry , Beilei He , Emmanuel Barillot , 2 ´ 1,3,4,5 2 Thierry Dubois , Philippe Hupe , Leanne de Koning * 1 Institut Curie, Paris, France, 2 Department of Translational Research, Institut Curie, Paris, France, 3 INSERM, U900, Paris, France, 4 Mines ParisTech, Fontainebleau, France, 5 CNRS UMR144, Paris, France Abstract Motivation: Reverse phase protein array (RPPA) is a powerful dot-blot technology that allows studying protein expression levels as well as post-translational modifications in a large number of samples simultaneously. Yet, correct interpretation of RPPA data has remained a major challenge for its broad-scale application and its translation into clinical research. Satisfying quantification tools are available to assess a relative protein expression level from a serial dilution curve. However, appropriate tools allowing the normalization of the data for external sources of variation are currently missing. Results: Here we propose a new method, called NormaCurve, that allows simultaneous quantification and normalization of RPPA data. For this, we modified the quantification method SuperCurve in order to include normalization for (i) background fluorescence, (ii) variation in the total amount of spotted protein and (iii) spatial bias on the arrays. Using a spike-in design with a purified protein, we test the capacity of different models to properly estimate normalized relative ex