Implementation of an Internalized Hemolysis Assessment Approach in Targeted Bottom-Up Proteomics for the Purpose of Blood Plasma Sample Quality Control
Anastasia V. Vakaryuk, Ivan O. Butenko, Natalia A. Kitsilovskaya, Alexey V. Kovalenko, Veronica D. Gremyacheva, Grigory L. Kozhemyakin, Angelina A. Lebedeva, Nikita M. Baraboshkin, Oleg V. Fedorov, Olga V. Kurilova, Konstantin S. Gorbunov, Irina V. Kolesnikova, Lyubov A. Davydova, Vadim M. GovorunAbstract
Mishandling of samples during blood plasma preparation may lead to biases in proteomic analysis and misinterpretation of data. An earlier study by Geyer et al. (2019) described proteins that may indicate poor plasma sample quality. Following this work, we have imitated a triangular strategy for biomarker discovery, established a targeted quantitative method for panels of hemolyzed and coagulated plasma sample markers proposed in the previous study, and validated it in our own model experiment. We have developed a scale for estimating the degree (rate) of hemolysis based on several protein markers and investigated the feasibility of using an internal control approach to detect hemolysis in targeted proteomic analysis. We propose including our plasma sample quality assessment and associated sample exclusion criteria into standard MRM analyses to provide more reliable data for proteomic research.