BookTesshi Yamada, Satoshi S. Nishizuka, Gordon B. Mills, Lance A. Liotta, editors.
Contents:
Intro
Reverse Phase Protein Array (RPPA) Global Workshop
Contents
1: RPPA: Origins, Transition to a Validated Clinical Research Tool, and Next Generations of the Technology
1.1 Opportunities and Challenges in Precision Medicine
1.2 Historical Origin of Reverse Phase Protein Microarray Technology
1.2.1 Protein Microarray Terminology
1.3 Application of RPPA Technology to Profile Individual Patient's Tumor Specimens 1.3.1 Near-Term Future Directions for RPPA (ISPY2): Discovery of Activated Kinase Signal Pathway Proteins Predicting Patient Response to New Molecular Therapy Agents
1.3.2 Side-Out Metastatic Breast Cancer Trial of Personalized Therapy
1.4 The Next Generation of RPPA Technology
1.4.1 Electronic RPPA
1.4.2 Solution Phase RPPA
1.5 Conclusions and Vision for the Future
References
2: Lysate Preparation for Reverse Phase Protein Arrays
2.1 Introduction
2.2 General Considerations
2.3 Protein Isolation from FFPE Tissues 2.4 Deparaffinization and Protein Isolation from Slide-Mounted FFPE Sections
2.4.1 Reagents
2.4.2 Equipment
2.4.3 Method
2.4.3.1 Sample Preparation
2.4.3.2 Deparaffinization
2.4.3.3 Protein Isolation
2.5 Deparaffinization and Protein Isolation from FFPE Sections Cut Directly in a Tube
2.5.1 Reagents
2.5.2 Equipment
2.5.3 Method
2.5.3.1 Sample Preparation
2.5.3.2 Deparaffinization
2.5.3.3 Protein Extraction
2.6 Lysate Preparation
2.7 Troubleshooting
References 3: High Precision RPPA: Concept, Features, and Application Performance of the Integrated Zeptosens Platform
3.1 Introduction
3.2 Platform Considerations
3.2.1 Required Sensitivity
3.2.2 RPPA Alternative Approaches and Differentiation
3.2.3 Detrimental Optical Effects
3.2.4 Sample Integrity and Deterioration
3.2.5 Microarray Layout and Printing
3.2.6 Antibodies and Assays
3.2.7 Data Analysis and Quantitation
3.2.8 Information Standardization
3.2.9 Quality Demands of Drug Development and Clinical Research
3.3 Realization - Implementation 3.3.1 Sample Preparation
3.3.2 Assay and Printing Buffers
3.3.3 Microarray Substrates
3.3.4 RPPA Printing
3.3.5 Microarray Layout
3.3.6 Assay Handling Hardware and Processing
3.3.7 Assay Architecture and Antibodies
3.3.8 Fluorescence Labels
3.3.9 Imaging via the ZeptoREADER
3.3.10 Image and Data Analysis
3.3.11 Quality Issues
3.4 Application Performance Obtainable from Zeptosens RPPA
3.5 Future Needs and Trends
References
4: Solid Pin Protein Array Printing Platforms
4.1 Introduction to Reverse Phase Protein Arrays 4.1.1 Workflow for Fabricating RPPA