A next generation setup for pre-fractionation of non-denatured proteins reveals diverse albumin proteoforms each carrying several post-translational modifications

Heidrun Rhode*, Petra Muckova, Rita Büchler, Sindy Wendler, Bärbel Tautkus, Michaela Vogel, Thomas Moore, Julian Grosskreutz, Andree Klemm, Mary Nabity

*Corresponding author for this work
4 Citations (Scopus)

Abstract

Proteomic biomarker search requires the greatest analytical reproducibility and detailed information on altered proteoforms. Our protein pre-fractionation applies orthogonal native chromatography and conserves important features of protein variants such as native molecular weight, charge and major glycans. Moreover, we maximized reproducibility of sample pre-fractionation and preparation before mass spectrometry by parallelization and automation. In blood plasma and cerebrospinal fluid (CSF), most proteins, including candidate biomarkers, distribute into a multitude of chromatographic clusters. Plasma albumin, for example, divides into 15-17 clusters. As an example of our technique, we analyzed these albumin clusters from healthy volunteers and from dogs and identified cluster-typical modification patterns. Renal disease further modifies these patterns. In human CSF, we found only a subset of proteoforms with fewer modifications than in plasma. We infer from this example that our method can be used to identify and characterize distinct proteoforms and, optionally, enrich them, thereby yielding the characteristics of proteoform-selective biomarkers.

Original languageEnglish
Article number11733
JournalScientific Reports
Volume9
Issue number1
ISSN2045-2322
DOIs
Publication statusPublished - 01.12.2019
Externally publishedYes

Research Areas and Centers

  • Centers: Center for Neuromuscular Diseases

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