Introducing plasma/serum glycodepletion for the targeted proteomics analysis of cytolysis biomarkers

A major class of clinical biomarkers is constituted of intracellular proteins which are leaking into the blood following ischemia, exposure to toxic xenobiotics or mechanical aggression. Their ectopic presence in plasma/serum is an indicator of tissue damage and raises a warning signal. These proteins, referred to as cytolysis biomarkers, are generally of cytoplasmic origin and as such, are devoid of glycosylation. In contrast, most plasma/serum proteins originate from the hepatic secretory pathway and are heavily glycosylated (at the exception of albumin). Recent advances in targeted proteomics have supported the parallelized evaluation of new blood biomarkers. However, these analytical methods must be combined with prefractionation strategies that reduce the complexity of plasma/serum matrix. In this article, we present the glycodepletion method, which reverses the hydrazide-based glycocapture concept to remove plasma/serum glycoproteins from plasma/serum matrix and facilitates the detection of cytolysis biomarkers. Glycodepletion was integrated to a targeted proteomics pipeline to evaluate 4 liver cytolysis biomarker candidates in the context of acetaminophen-induced acute hepatitis. [Display omitted] •Targeted proteomics assays are increasingly used for the parallelized evaluation of blood biomarker candidates.•Up-front fractionation strategies are required to reduce the protein complexity of plasma/serum samples.•We describe glycodepletion as a novel fractionation procedure for the detection of cytolysis biomarkers (non-glycosylated proteins).•Serum glycodepletion and targeted proteomics were used to evaluate 4 liver cytolysis biomarker candidates in clinical samples.