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Affinity Membranes and Monoliths for Protein Purification

机译：亲和膜和整体结构用于蛋白质纯化

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Affinity capture represents an important step in downstream processing of proteins and it is conventionally performed through a chromatographic process. The performance of this step highly depends on the type of matrix employed. In particular, resin beads and convective materials, such as membranes and monoliths, are the commonly available supports. The present work deals with non-competitive binding of bovine serum albumin (BSA) on different chromatographic media functionalized with Cibacron Blue F3GA (CB). The aim is to set up the development of the purification process starting from the lab-scale characterization of a commercially available CB resin, regenerated cellulose membranes and polymeric monoliths, functionalized with CB to identify the best option. The performance of the three different chromatographic media is evaluated in terms of BSA binding capacity and productivity. The experimental investigation shows promising results for regenerated cellulose membranes and monoliths, whose performance are comparable with those of the packed column tested. It was demonstrated that the capacity of convective stationary phases does not depend on flow rate, in the range investigated, and that the productivity that can be achieved with membranes is 10 to 20 times higher depending on the initial BSA concentration value, and with monoliths it is approximately twice that of beads, at the same superficial velocity.
机译：亲和捕获代表了蛋白质下游加工中的重要步骤，并且通常通过色谱过程进行。此步骤的性能高度取决于所用矩阵的类型。特别地，树脂珠和对流材料，例如膜和整料，是通常可获得的载体。目前的工作涉及牛血清白蛋白（BSA）在不同层析介质上的非竞争性结合，所述层析介质具有Cibacron Blue F3GA（CB）功能。目的是从实验室规模表征市售CB树脂，再生纤维素膜和聚合物整料开始，以CB进行功能化以鉴定最佳选择，从而建立纯化工艺的开发。根据BSA结合能力和生产率评估了三种不同色谱介质的性能。实验研究表明，再生纤维素膜和整体填料的性能令人鼓舞，其性能可与测试的填充柱相媲美。结果表明，在所研究的范围内，对流固定相的容量不取决于流速，取决于初始BSA浓度值和整体式填料，使用膜可以达到的生产率高出10到20倍。在相同的表观速度下，大约是珠子的两倍。

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