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CASE STUDY: Biomarker Quantitation in Human Plasma by Multiple Reaction Monitoring (MRM)

Discovering new protein biomarkers is critical in scientific understanding of disease pathway mechanisms. Novel protein biomarkers provide the basis for earlier disease diagnosis, more accurate prognosis of disease aggression, and better prediction of patient response to therapy. Researchers and clinicians are actively pursuing toolsets for the discovery and verification of new protein biomarker candidates. To deliver a quantitative determination of targeted biomarkers, Protea’s interdisciplinary team developed workflows utilizing a range of sample preparation methods and techniques, along with a variety of analytical approaches.

One of the most effective tools Protea offers is the absolute quantitation of targeted protein by mass spectrometry-based multiple reaction monitoring (MRM) [1-3]. This technique is quickly becoming the standard for protein biomarker verification. MRM combined with a stable isotope-labeled internal standard provides strategic advantages compared to the traditional immunoassay platforms. This combination delivers the ability to multiplex and provides improved accuracy, precision, and specificity. The method development timeline is also faster and more cost-effective.

In this investigation, nine signature peptides were used for quantitation of the original seven proteins fortified into female human plasma. The accuracy, precision, sensitivity, and linearity of targeted protein quantitation by MRM were evaluated. We confidently quantified two human proteins and five proteins from other species previously detected at low levels.

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