MSACL 2019 US Abstract

Improving the Sensitivity and Accuracy of Quantitative Protein Biomeasure Mass Spectrometry Through the Use of a SRM Transition Summing Technique

Jay S. Johnson, Joe Palandra, Jason M. Walsh, Katherine Wright, Hendrik Neubert
Biomedicine Design, Pfizer Inc, Andover, MA

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Jay Johnson (Presenter) Pfizer Worldwide Research and Development

Presenter Bio: Jay received his B.S. in Chemistry from The George Washington University in 2006 and his M.S. in Analytical Chemistry from Northeastern University in 2011. For the first ten years of his career he worked at Waters Corporation where he acquired extensive knowledge and expertise in low flow chromatography and played a key role in the development and support of the ionKey/MS system in collaborator, customer, and demo labs worldwide.
Jay joined the Hendrik Neubert’s Biomeasures group at Pfizer in February of 2017 with the goal of honing his skill in large molecule bioanalytical sample preparation techniques and developing workflows to increase instrument sensitivity and speed for online immunoaffinity nanoflow LC/MS approaches. To that end, he has supported a variety of projects including protein target quantitation in a variety matrices and protein synthesis measurements.

Relevant Financial Disclosures (within past 24 months) Honorarium/Expenses Pfizer Stock/Bonds Pfizer, Waters Corp Salary Pfizer Discussing Products? NO, presenter will NOT present educational content related to products/services of the ineligible company(ies) listed above. Conflict mitigation NOT REQUIRED.

The desire to reach ever diminishing lower limits of quantitation (LLOQ) and probe changes in the lowest endogenously expressed proteins has led to enormous progress in LC-MS/MS instrumentation and technique. To maximize sensitivity and reduce noise we often combine immunoaffinity enrichment with nanoflow LC-MS/MS. However, situations arise where these advances are still insufficient in attaining the needed LLOQ. Here we examine the successful deployment of collecting transitions in a different way, utilizing a SRM transition summing technique, to improve sensitivity, reduce LLOQ, and deliver more accurate quantitative values for the biomeasure of Nerve Growth Factor in serum, inflammatory cytokines in atopic dermatitis skin biopsies, and to determine the protein synthesis rate of a cytokine receptor for incorporation in mechanistic PK/PD models to inform translational pharmacology.