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用于生物分离的具有高蛋白结合能力的非织造离子交换膜。

Nonwoven Ion-Exchange Membranes with High Protein Binding Capacity for Bioseparations.

作者信息

Lemma Solomon Mengistu, Boi Cristiana, Carbonell Ruben G

机构信息

Golden LEAF Biomanufacturing Training and Education Center (BTEC), North Carolina State University, 850 Oval Dr, Raleigh, NC 27695-7905, USA.

DICAM, Alma Mater Studiorum-Università di Bologna, via Terracini 34, 40128 Bologna, Italy.

出版信息

Membranes (Basel). 2021 Mar 6;11(3):181. doi: 10.3390/membranes11030181.

Abstract

This study presents the preparation and characterization of UV-grafted polybutylene terepthalate (PBT) ion exchange nonwoven membranes for chromatographic purification of biomolecules. The PBT nonwoven was functionalized with sulfonate and secondary amine for cation and anion exchange (CEX and AEX), respectively. The anion exchange membrane showed an equilibrium static binding capacity of 1300 mg BSA/g of membrane, while the cationic membranes achieved a maximum equilibrium binding capacity of over 700 mg hIgG/g of membrane. The CEX and AEX membranes resulted in dynamic binding capacities under flow conditions, with a residence time of 0.1 min, of 200 mg hIgG/mL of membrane and 55 mg BSA/mL of membrane, respectively. The selectivity of the PBT-CEX membranes was demonstrated by purifying antibodies and antibody fragments (hIgG and scFv) from CHO cell culture supernatants in a bind-an-elute mode. The purity of the eluted samples exceeded 97%, with good log removal values (LRV) for both host cell proteins (HCPs) and DNA. The PBT-AEX nonwoven membranes exhibited a DNA LRV of 2.6 from hIgG solutions in a flow-through mode with little loss of product. These results indicate that these membranes have significant potential for use in downstream purification of biologics.

摘要

本研究介绍了用于生物分子色谱纯化的紫外线接枝聚对苯二甲酸丁二醇酯(PBT)离子交换非织造膜的制备与表征。PBT非织造布分别用磺酸盐和仲胺进行功能化处理,以实现阳离子交换(CEX)和阴离子交换(AEX)。阴离子交换膜的平衡静态结合容量为1300 mg BSA/g膜,而阳离子膜的最大平衡结合容量超过700 mg hIgG/g膜。在流动条件下,停留时间为0.1分钟时,CEX和AEX膜的动态结合容量分别为200 mg hIgG/mL膜和55 mg BSA/mL膜。通过以结合-洗脱模式从CHO细胞培养上清液中纯化抗体和抗体片段(hIgG和scFv),证明了PBT-CEX膜的选择性。洗脱样品的纯度超过97%,对宿主细胞蛋白(HCPs)和DNA均具有良好的对数去除值(LRV)。PBT-AEX非织造膜在流通模式下对hIgG溶液的DNA LRV为2.6,且产物损失很小。这些结果表明,这些膜在生物制品下游纯化中具有巨大的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4e76/8001514/87d7b5340f8f/membranes-11-00181-g001.jpg

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