Precise quantitation of urinary proteins for biomarker assessment

10^th European Nephrology Conference

October 24-26, 2016 Rome, Italy

Christoph Borchers

University of Victoria, Canada

Posters & Accepted Abstracts: J Nephrol Ther

Abstract :

Quantitative MS is increasingly being performed in the proteomics field to address biological- and medical-related queries. While this can be achieved with a relative or absolute technique, it is the latter that is more informative and better suited for protein biomarker evaluation of pre-clinical and clinical samples. Such absolute quantitative techniques determine endogenous protein concentrations and require the use of stable isotope-labeled standards (SIS), along with a targeted form of MS detection (typically MRM), to be employed within a bottom-up analytical workflow. Using that approach on non-invasively collected urine samples, we have systematically advanced the methodology of extended breadth and depth of protein quantitation. The final method enabled the reproducible quantitation of 136 proteins spanning >5 orders of magnitude in concentration (from 8.6 �¼g/mL to 25 pg/mL). The lower complexity of the urine matrix enabled lower-abundance plasma proteins (e.g., osteopontin) to be detected without prefractionation, which is a significant advantage of using urine for protein biomarker screening. This quantitative method was then applied to the analysis of 53 time-course urine samples to estimate excretion rates of proteins from the kidneys. This presentation will discuss the method development, provide an overview of its application and its extension to kits for the research community.

Biography :

Email: christoph@proteincentre.com