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Gewählte Publikation:

Farcet, JB; Kosma, P.
(2021): Determination of modification degree of polysialylated therapeutic proteins using 1H-NMR spectroscopy
INT J BIOL MACROMOL. 2021; 185: 1015-1021. FullText FullText_BOKU

Abstract:
Water soluble polymers and their derivatives bound to proteins can dramatically favor the biological activity of new drugs and vaccines. Quantification of the modification degree of the protein is crucial during the development and licensing phase and later in order to monitor the industrial production process and to match product specification. In this work, we describe an innovative way to measure directly the modification degree of polysialylated proteins using proton NMR (Nuclear Magnetic Resonance) spectroscopy. Following a calibration step, the modification degree can be easily deduced by the integration ratio of a separate signal from the polymer and selected signals from the protein. In fact, the upfield-shifted signals of methyl groups from Valine, Leucine and Isoleucine can be used as an internal calibration reference for the integration. In this paper recombinant factor VIII (rFVIII) and recombinant factor IX (rFIX) proteins modified by polysialic acid (PSA) are used to illustrate the accuracy, reproducibility and ease of the method that may replace or complement wet-chemistry approaches.
Autor*innen der BOKU Wien:
Farcet Jean-Baptiste
Kosma Paul
BOKU Gendermonitor:


Find related publications in this database (Keywords)
Modification degree
Quantitative NMR
Polysialic acid
Resorcinol assay


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