Polymeric short-fiber chromatographic supports for downstream processing of biomolecules

Anirudh Singh , Neville G. Pinto
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引用次数: 21

Abstract

Fiber chromatography is prominent among the novel systems being developed for downstream processing. In this research, a polymer based anion-exchange support with fiber geometry has been developed. The fiber support has an asymmetric structure, with a dense skin and a porous substructure. Equilibrium adsorption capacities of this support for bovine serum albumin and β-lactoglobulin were measured. Isotherms were obtained at different pH and salt concentrations, and were found to be Langmuirian. It was also found that ion-exchange is the dominant adsorption mechanism, and, due to a large internal surface area, the support has a high adsorption capacity. The fiber support was packed in a bed, and tested for its applicability in the isolation of proteins from dilute solutions. The fiber column is shown to have a significantly lower pressure drop than columns packed with commercial, spherical supports. The frontal throughout for this column was evaluated at different conditions. The throughput was found to increase with flow rate and protein concentration, but the yield dropped with an increase in flow rate. However, the throughput and yield were high when a high concentration of protein was loaded at a low flow rate and washed at a very high flow rate.

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聚合物短纤维色谱支持下游处理的生物分子
纤维色谱法在正在开发的用于下游加工的新系统中是突出的。在本研究中,开发了一种基于聚合物的具有纤维几何形状的阴离子交换载体。所述纤维支架具有不对称结构,具有致密的表皮和多孔的下层结构。测定了该载体对牛血清白蛋白和β-乳球蛋白的平衡吸附能力。在不同的pH和盐浓度下得到等温线,发现等温线为Langmuirian。离子交换是主要的吸附机制,并且由于支架内表面积大,具有较高的吸附容量。将纤维支架包装在床上,并测试其在从稀溶液中分离蛋白质方面的适用性。结果表明,纤维柱的压降明显低于装有商业球形支撑的柱。在不同的条件下评估了这一列的正面贯穿。通量随流速和蛋白质浓度的增加而增加,但产量随流速的增加而下降。然而,当以低流速装载高浓度蛋白质并以非常高的流速洗涤时,吞吐量和产量很高。
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