Mass Spectrometry of Biotherapeutics – Developments in Dextran MS Analysis and Identification of Interferences in Metabolic Assays
Research seminar abstract
The use of materials for biomedical applications must be evaluated for their physiological benefits, but also for their degradation byproducts and potential interferences. The focus of this presentation will relate to the measurement of dextran and the identification of interfering species in common metabolic assays using mass spectrometry. Dextran is a glucose-derived polysaccharide1 with important applications in the food industry2, chromatographic separations1, and as a biotherapeutic material in the form of a blood plasma expander (BPE) and NO-releasing dextran.3 As a BPE and NO-releasing polysaccharide, it is important to track its degradation over time after administration to patients to determine appropriate dosage/molecular weight of dextran. To track dextran degradation, a one-pot derivatization of dextran with ethylenediamine was developed4,5 to encourage ionization by multiply-charging dextran, a feature not previously observed with the polysaccharide. This allowed derivatized dextran to get a maximum of four-protonations/charges, shifting the ion peaks toward a lower m/z range.5 Secondly, the effect of potential interferences in the commonly used resazurin and MTT in vitro cytotoxicity assays were studied. The standard assay protocols were performed with the addition of mass spectrometric analysis of the resulting media to determine the extent to which the potential interferences affect the assays’ results.6 The results from this study indicate that more in-depth control studies must be performed to obtain the most accurate data from these assays. Collectively, these studies provide insight into mass spectrometric techniques for dextran analysis and potential interferences in commonly used cytotoxicity assays.
(1) Naessens, M.; Cerdobbel, A.; Soetaert, W.; Vandamme, E.J. J. Chem. Technol. Biotechnol. 2005, 80(8), 845
(2) Katina, K.; Maina, N.H.; Juvonen, R.; Flander, L.; Johansson, L.; Virkki, L.; Tenkanen, M. Food Microbiol. 2009, 26(7), 734.
(3) Damodaran, V.B.; Place, L.W.; Kipper, M.J.; Reynolds, M.M. J. Mater. Chem. 2012, 22, 23038.
(4) Nakamura, J.; Nakajima, N.; Matsumura, K.; Hyon, S.H. Anticancer Res. 2010, 30, 903.
(5) Tapia, J.B.; Hibbard, H.A.J.; Reynolds, M.M. J. Amer. Soc. Mass Spectrom. 2017, 28(10), 2201.
(6) Neufeld, B.H.; Tapia, J.B.; Lutzke, A.; Reynolds, M.M. Anal. Chem. 2018, Manuscript submitted for publication
Speaker: Jesus Tapia
Speaker Institution: Colorado State University
Event Date: 01-17-2018
Event Time: 4:00 PM
Event Location: Chemistry A101
Mixer Time: 3:45 PM
Mixer Location: Chemistry B101E
Host: M. Reynolds