Rapid determination of sphingosine 1-phosphate association with carrier molecules by flow-induced dispersion analysis to predict sepsis outcome
Isabelle Seidita,
Anke Ziegler,
Auron Qalaj,
Martin Sebastian Winkler,
Axel Nierhaus,
Stefan Kluge,
Bodo Levkau,
Markus H. Gräler
Affiliations
Isabelle Seidita
Department of Anesthesiology and Intensive Care Medicine, Center for Molecular Biomedicine (CMB), Jena University Hospital, Hans-Knöll-Straße 2, D-07745 Jena, Germany
Anke Ziegler
Department of Anesthesiology and Intensive Care Medicine, Center for Molecular Biomedicine (CMB), Jena University Hospital, Hans-Knöll-Straße 2, D-07745 Jena, Germany
Auron Qalaj
Department of Anesthesiology and Intensive Care Medicine, Center for Molecular Biomedicine (CMB), Jena University Hospital, Hans-Knöll-Straße 2, D-07745 Jena, Germany
Martin Sebastian Winkler
Department of Anesthesiology, Emergency and Intensive Care Medicine, University of Göttingen, Robert-Koch-Straße 40, 37075 Göttingen, Germany
Axel Nierhaus
Department of Intensive Care, Universitätsklinikum Eppendorf, Martinistraße 52, 20246 Hamburg, Germany
Stefan Kluge
Department of Intensive Care, Universitätsklinikum Eppendorf, Martinistraße 52, 20246 Hamburg, Germany
Bodo Levkau
Institute of Molecular Medicine III, Heinrich Heine University, Universitätsstraße 1, 40225 Düsseldorf, Germany
Markus H. Gräler
Department of Anesthesiology and Intensive Care Medicine, Center for Molecular Biomedicine (CMB), Jena University Hospital, Hans-Knöll-Straße 2, D-07745 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07740 Jena, Germany; Corresponding author
Summary: Flow-induced dispersion analysis (FIDA) was used to investigate the association of fluorescein isothiocyanate-labeled signaling lipid sphingosine 1-phosphate (S1P) with its carrier molecules human serum albumin (HSA) and high-density lipoprotein (HDL). Associations were measured in plasma samples of patients after surgery, with sepsis or septic shock. All patients demonstrated a significant shift between the carrier binding: decrease of S1P bound to HSA with a concomitant increase of S1P bound to HDL. The molecular sizes of binding complexes correlated well with the relative amounts of S1P bound to HSA and HDL detected by liquid chromatography-tandem mass spectrometry. Very low complex formation of S1P with HDL was observed in several septic shock patients and correlated with the need for mechanical ventilation and intensive care unit (ICU) mortality. Determination of S1P binding to HSA and HDL by FIDA could therefore be useful in the clinical setting to predict disease progression, severity, and outcome.
