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Composite cryogel with immobilized concanavalin A for affinity chromatography of glycoproteins
Authors:Solmaz Hajizadeh  Harald Kirsebom  Andre Leistner  Bo Mattiasson
Institution:1. Department of Biotechnology, Lund University, , Lund, Sweden;2. Polymerics GmbH, , Berlin, Germany
Abstract:Composite cryogels containing porous adsorbent particles were prepared under cryogelation conditions. The composites with immobilized concanavalin A (Con A) were used for capturing glycoproteins. Adsorbent particles were introduced into the structure in order to improve the capacity and to facilitate the handling of the particles. The monolithic composite cryogels were produced from suspensions of polyvinyl alcohol particles and porous adsorbent particles and cross‐linked under acidic conditions at sub‐zero temperature. The cryogels were epoxy activated and Con A was immobilized as an affinity ligand. Binding and elution of horseradish peroxidase (HRP) was studied in batch experiment and in a chromatographic setup. Increasing adsorbent concentration in composite cryogels will increase ligand density, which therefore enhances the amount of bound HRP from 0.98 till 2.9 (milligram enzyme per milliliter of gel) in the chromatographic system. The material was evaluated in 10 cycles for binding and elution of HRP.
Keywords:Adsorbent polymer  Affinity chromatography  Composite  Con A  Cryogel  Glycoprotein
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