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Application of lectin immobilized on polyHIPE monoliths for bioprocess monitoring of glycosylated proteins.
Stantic, Metka; Guncar, Gregor; Kuzman, Drago; Mravljak, Rok; Cvijic, Tamara; Podgornik, Ales.
Afiliación
  • Stantic M; Faculty for Chemistry and Chemical Technology, University of Ljubljana, Vecna pot 113, 1000 Ljubljana, Slovenia.
  • Guncar G; Faculty for Chemistry and Chemical Technology, University of Ljubljana, Vecna pot 113, 1000 Ljubljana, Slovenia.
  • Kuzman D; Technical development biosimilars, Global drug development, Novartis, Kolodvorska 27, 1234 Menges, Slovenia.
  • Mravljak R; Faculty for Chemistry and Chemical Technology, University of Ljubljana, Vecna pot 113, 1000 Ljubljana, Slovenia.
  • Cvijic T; Technical development biosimilars, Global drug development, Novartis, Kolodvorska 27, 1234 Menges, Slovenia.
  • Podgornik A; Faculty for Chemistry and Chemical Technology, University of Ljubljana, Vecna pot 113, 1000 Ljubljana, Slovenia; COBIK, Tovarniska 26, 5270 Ajdovscina, Slovenia. Electronic address: ales.podgornik@fkkt.uni-lj.si.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1174: 122731, 2021 Jun 01.
Article en En | MEDLINE | ID: mdl-33971517
In-process monitoring of glycosylated protein concentration becomes very important with the introduction of perfusion bioprocesses. Affinity chromatography based on lectins allows selective monitoring when carbohydrates are accessible on the protein surface. In this work, we immobilized lectin on polyHIPE type of monoliths and implemented it for bioprocess monitoring. A spacer was introduced to lectin, which increased binding kinetics toward Fc-fusion protein, demonstrated by bio-layer interferometry. Furthermore, complete desorption using 0.25 M galactose was shown. 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 four 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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Glicoproteínas / Cromatografía de Afinidad / Proteínas Inmovilizadas / Lectinas Límite: Animals Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2021 Tipo del documento: Article País de afiliación: Eslovenia Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Glicoproteínas / Cromatografía de Afinidad / Proteínas Inmovilizadas / Lectinas Límite: Animals Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2021 Tipo del documento: Article País de afiliación: Eslovenia Pais de publicación: Países Bajos