Broadcast Date: February 25th, 2015
A significant challenge to standardization in liquid chromatography-multiple reaction monitoring (LC-MRM) workflows is the variability caused by the multistep, highly manual nature of sample preparation. If MRM-based protein quantification is to enjoy widespread use for disease biomarker assessment studies—and ultimately secure acceptance for clinical analysis—the technique must be standardized to facilitate precise protein quantification. To address the issues of throughput and variability related to complex manual workflows, automation platforms are being developed to serve as the foundation for sample preparation in LC-MRM assays.
A typical sample preparation workflow for LC-MRM measurements of human plasma proteins involves several manual manipulations including protein solubilization/denaturation, reduction, alkylation, trypsinization, and subsequent solid-phase extraction (SPE). These manual tasks can be performed on the AssayMAP Bravo Platform, a state-of-the-art liquid handler equipped with an AssayMAP liquid-handling head. Tasso Miliotis, Ph.D., associate principal scientist at AstraZeneca, will discuss how his team has incorporated AssayMAP into their workflow and describe the results they’ve achieved.
Steve Murphy Ph.D., director of Agilent AssayMAP applications development, will provide some additional detail about the capabilities of the AssayMAP platform for this application.
Who Should Attend
- R & D scientists
- Proteomics scientists
- Bioanalytical scientists
- Researchers interested in protein quantification
- Biomarker assay developers
You Will Learn
- How LC-MRM is used for absolute quantification of proteins using stable isotope-labeled standards.
- Challenges in the workflow, and potential solutions.
- Important things to consider when automating the workflow.
- How the workflow can be improved using the AssayMAP Bravo Platform.
Produced with support from:
Tasso Miliotis, Ph.D.
Associate Principal Scientist,
Steve Murphy, Ph.D.
Director of AssayMAP Applications Development