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Reverse phase protein arrays in acute leukemia: investigative and methodological challenges
Fieke W Hoff1, Terzah M Horton2, Steven M Kornblau3
1Department of Internal Medicine, The University of Texas Southwestern Medical Center, TX, USA.
Expert Review of Proteomics
|December 29, 2021
Summary
Reverse phase protein arrays (RPPA) are effective for studying protein changes in acute leukemia. This review covers technical and analytical issues for developing RPPAs, emphasizing sample preparation and antibody validation for accurate proteomic analysis.
Area of Science:
- Proteomics
- Molecular Biology
- Oncology
Background:
- Acute leukemia arises from mutations altering cell growth and proliferation.
- Protein changes are central to leukemia development.
- Proteomics offers a method to study these protein alterations.
Purpose of the Study:
- To review the technical, methodological, and analytical aspects of developing Reverse Phase Protein Arrays (RPPA) for acute leukemia.
- To guide the successful implementation of RPPA technology in leukemia research.
Main Methods:
- Utilizing high-throughput Reverse Phase Protein Array (RPPA) technology for protein expression assessment.
- Analyzing protein samples from clinical trials.
- Discussing sample preparation, preservation, and antibody validation.
Main Results:
- Freshly collected blood or bone marrow samples yield representative protein lysates.
- Minimal impact of sample shipment variables on protein expression.
- CellSave preservation tubes are recommended for post-chemotherapy samples.
- Cell lines are not representative of patient proteomic patterns.
Conclusions:
- Standardized antibody validation and systematic biological approaches are crucial for RPPA development in leukemia.
- RPPA facilitates the classification of patients into risk groups based on protein expression patterns.
- Accurate proteomic analysis using RPPA can inform personalized treatment strategies.
