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Development and characterization of methacrylate-based hydrazide monoliths for oriented immobilization of antibodies
Authors:P Brne  Y-P Lim  A Podgornik  M Barut  B Pihlar  A Štrancar
Institution:1. BIA Separations d.o.o., Teslova 30, 1000 Ljubljana, Slovenia;2. ProThera Biologics, LLC, 551 Warren Avenue, East Providence, RI 02914, USA;3. Faculty of Chemistry and Chemical Technology, Ašker?eva cesta 5, 1000 Ljubljana, Slovenia
Abstract:Convective interaction media (CIM; BIA Separations) monoliths are attractive stationary phases for use in affinity chromatography because they enable fast affinity binding, which is a consequence of convectively enhanced mass transport. This work focuses on the development of novel CIM hydrazide (HZ) monoliths for the oriented immobilization of antibodies. Adipic acid dihydrazide (AADH) was covalently bound to CIM epoxy monoliths to gain hydrazide groups on the monolith surface. Two different antibodies were afterwards immobilized to hydrazide functionalized monolithic columns and prepared columns were tested for their selectivity. One column was further tested for the dynamic binding capacity.
Keywords:CIM  Convective interaction media  Monolith  Affinity chromatography  Hydrazide  Antibody immobilization
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