Stančič, Metka, Cvijic, Tamara, Kuzman, Drago, Mravljak, Rok, Gunčar, Gregor and Podgornik, Aleš (2021) Application of lectin immobilized on polyHIPE monoliths for bioprocess monitoring of Fc-fusion surface glycosylated protein. Journal of chromatography B : biomedical sciences and applications.

Abstract

In-process monitoring of glycosylated protein concentration becomes very important with introduction of perfusion bioprocesses. Affinity chromatography based on lectines allows selective monitoring of surface glycosylated proteins. In this work we immobilized lectine on polyHIPE type of monoliths and implemented it for bioprocess monitoring. Spacer was introduced to lectine, which increased binding kinetics toward fusion protein, demonstrated by biolayer interferometry. Furthermore, complete desorption using 0.25 M galactose was demonstrated. Affinity column exhibited linearity in the range between 0.5 and 8 mg/ml and flow-unaffected binding for the flow-rates between 0.5 and 8 ml/min. Long-term stability over at least 4 months period was demonstrated. No unspecific binding of culture media components, including host cell proteins and DNA, was detected. Results obtained by affinity column matched concentration values obtained by a reference method.

Item Type:	Article
Keywords:	lectin chromatography, polyHIPE monolith, glycosylated proteins, perfusion bioprocess monitoring
Date Deposited:	01 Jun 2021 00:45
Last Modified:	01 Jun 2021 00:45
URI:	https://oak.novartis.com/id/eprint/44231