Proteomics: from hypothesis to quantitative assay on a single platform. Guidelines for developing MRM assays using
1Lilly Corporate Center, Drop Code GL54, Indianapolis, IN 46285, USA. bomiehan@lilly.com
Briefings in Functional Genomics & Proteomics
|June 27, 2008
Summary
Developing targeted protein assays using multiple reaction monitoring (MRM) on ion trap mass spectrometers accelerates biomarker validation. This approach streamlines the transition from proteomic discovery to clinical application, enhancing disease and drug response analysis.
Area of Science:
- Proteomics and Mass Spectrometry
- Biomarker Discovery and Validation
- Clinical Assay Development
Background:
- High-throughput HPLC-mass spectrometry (HPLC-MS) is crucial for profiling biological samples to identify potential protein biomarkers for disease, drug efficacy, and toxicity.
- Translating proteomic profiling discoveries into clinical applications is a significant bottleneck, necessitating the development of more sensitive and selective assays.
- Multiple reaction monitoring (MRM) assays offer enhanced sensitivity, specificity, and quantitative capabilities, making them suitable for biomarker validation.
Purpose of the Study:
- To provide guidelines for the rapid development of multiple reaction monitoring (MRM) assays.
- To enable the use of the same mass spectrometry platform (ion traps) for both initial proteomic profiling and subsequent MRM assay development.
- To review methodological and analytical validation considerations for MRM assays in biomarker development.
Main Methods:
- Utilizing ion trap mass spectrometry platforms, traditionally used for high-throughput profiling, for the development of MRM assays.
- Adapting analytical validation guidelines from immunological assays for mass spectrometry-based biomarker assays.
- Focusing on a seamless transition from hypothesis generation via profiling to validation via MRM.
Main Results:
- Demonstrates a method for rapid MRM assay development on ion trap instruments.
- Outlines considerations for methodological and analytical validation applicable across different mass spectrometry platforms.
- Facilitates a faster pathway from proteomic discovery to validated clinical assays.
Conclusions:
- Developing MRM assays on the same platform used for proteomic profiling (ion traps) accelerates the biomarker translation process.
- The presented guidelines and validation considerations support the efficient development of sensitive and specific protein assays for clinical use.
- This approach helps overcome the bottleneck in translating proteomic discoveries into tangible clinical applications.
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