Abstract
Reverse Phase Protein Arrays (RPPA) represent a very promising sensitive and precise high-throughput technology for the quantitative measurement of hundreds of signaling proteins in biological and clinical samples. This array format allows quantification of one protein or phosphoprotein in multiple samples under the same experimental conditions at the same time. Moreover, it is suited for signal transduction profiling of small numbers of cultured cells or cells isolated from human biopsies, including formalin fixed and paraffin embedded (FFPE) tissues. Owing to the much easier sample preparation, as compared to mass spectrometry based technologies, and the extraordinary sensitivity for the detection of low-abundance signaling proteins over a large linear range, RPPA have the potential for characterization of deregulated interconnecting protein pathways and networks in limited amounts of sample material in clinical routine settings. Current aspects of RPPA technology, including dilution curves, spotting, controls, signal detection, antibody validation, and calculation of protein levels are addressed.
Highlights
The aim of future medicine is personalized treatment which is often characterized as providing the right patient with the right drug at the right dose at the right time
The importance of detecting phosphoproteins in tumor samples is illustrated by a recent study that revealed that patients with HER2 negative breast cancers (IHC/fluorescence in situ hybridization (FISH)) express a phosphorylated form of HER2 [6]
Reverse Phase Protein Arrays (RPPA) was one of the methods to determine if multi-omic molecular profiling based treatment improves the clinical course of patients with metastatic breast cancer measured by growth of modulation index (GMI)
Summary
The aim of future medicine is personalized treatment which is often characterized as providing the right patient with the right drug at the right dose at the right time. The importance of detecting phosphoproteins in tumor samples is illustrated by a recent study that revealed that patients with HER2 negative breast cancers (IHC/FISH) express a phosphorylated form of HER2 [6]. These phospho-HER2 positive patients may benefit from anti-HER2 treatments in addition to patients showing HER2 gene amplification. RPPA has proven to be useful for efficient tissue protein quantification and for analysis of signaling cascades even on the basis of very low amounts of tumor sample (biopsy), enabling quantification of biomarkers in tumors in neoadjuvant settings and at early time points [13,14,15]. We will highlight the clinical use of RPPA and discuss in detail recent developments for optimizing the RPPA procedure
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
