Abstract
Membrane based ion-exchange (IEX) and hydrophobic interaction chromatography (HIC) for protein purification is often used to remove impurities and aggregates operated under the flow-through mode. IEX and HIC are also limited by capacity and recovery when operated under bind-and-elute mode for the fractionation of proteins. Electrospun nanofibrous membrane is characterized by its high surface area to volume ratio and high permeability. Here tertiary amine ligands are grafted onto the electrospun polysulfone (PSf) and polyacrylonitrile (PAN) membrane substrates using UV-initiated polymerization. Static and dynamic binding capacities for model protein bovine serum albumin (BSA) were determined under appropriate bind and elute buffer conditions. Static and dynamic binding capacities in the order of ~100 mg/mL were obtained for the functionalized electrospun PAN membranes whereas these values reached ~200 mg/mL for the functionalized electrospun PSf membranes. Protein recovery of over 96% was obtained for PAN-based membranes. However, it is only 56% for PSf-based membranes. Our work indicates that surface modification of electrospun membranes by grafting polymeric ligands can enhance protein adsorption due to increased surface area-to-volume ratio.
Highlights
IntroductionThe production of protein-based human therapeutics such as monoclonal antibodies (mAbs) and
The global market of biologics, protein therapeutics is growing rapidly [1,2].The production of protein-based human therapeutics such as monoclonal antibodies andFc-fusion proteins involves cultivating mammalian cells such as Chinese hamster ovary cells (CHO)in complex cell culture suspension [3]
There has been significant work [8,9,10,11,12,13,14,17,18,19] on surface modification by grafting polymers on the flat sheet membrane substrates using atom-transfer radical polymerization (ATRP) and UV-initiated polymerizations. These results show that there exists an optimal ligand chain density and chain length for maximizing protein binding capacities for IEX, affinity and hydrophobic interaction chromatography (HIC) membrane chromatography [19]
Summary
The production of protein-based human therapeutics such as monoclonal antibodies (mAbs) and. In complex cell culture suspension [3]. The desired protein therapeutics are typically secreted by the cells into the suspension media. The rapid advancement in upstream cell culture operations has led to a significant increase in product titers. This high-level of productivity is accompanied by establishing much higher cell density [4,5] which places a much larger burden on the traditional downstream clarification and purification operations. Downstream processing becomes the bottleneck in the production of protein therapeutics and contributes significantly to the production cost [6]. High capacity and high recovery downstream purification unit operations are essential for the cost-effective purification of biologics
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