Mertz Michael, Golombek Florian, Boye Susanne, Moreno Silvia, Castiglione Kathrin
Lehrstuhl für Bioverfahrenstechnik, Friedrich-Alexander University Erlangen-Nürnberg, Paul-Gordan-Straße 3, 91052 Erlangen, Germany.
Leibniz-Institut für Polymerforschung Dresden e.V., Hohe Straße 6, 01069 Dresden, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jan 1;1162:122459. doi: 10.1016/j.jchromb.2020.122459. Epub 2020 Nov 28.
Artificial vesicles made of block copolymers, so-called polymersomes, represent a versatile chassis for the creation of functionalized nanocompartments with a wide range of biotechnological applications. The specific application depends on the biomolecules - usually proteins - that are positioned in the interior, in the membrane or on the surface of the vesicles. However, not all added proteins are integrated into the vesicles during the usual manufacturing processes of polymersomes. Excess proteins must therefore be removed. The separation techniques currently used for this, however, are associated with decisive disadvantages, such as damaged vesicles, long process times, or small sample volumes that can be processed. To overcome these drawbacks, we investigated the applicability of Capto™ Core 700 resin for polymersome purification. Polymersomes were not damaged or otherwise affected by passage through the column verified by hollow fiber flow field flow fractionation technique. Using three proteins with divergent physico-chemical properties as examples, it was demonstrated that different types of unentrapped proteins were efficiently removed from polymersome dispersions. The dynamic binding capacities in the presence of polymersomes varied between 9.5 and 16.5 mg per mL resin for the proteins applied. The technique can be used for small and large sample volumes alike. In addition, it can be used without special laboratory equipment. This adds a new and easy-to-use purification method for polymer vesicles to the repertoire that will also facilitate the large-scale production of functionalized polymersomes.
由嵌段共聚物制成的人工囊泡，即所谓的聚合物囊泡，是用于创建具有广泛生物技术应用的功能化纳米隔室的通用框架。具体应用取决于位于囊泡内部、膜上或表面的生物分子——通常是蛋白质。然而，在聚合物囊泡的常规制造过程中，并非所有添加的蛋白质都能整合到囊泡中。因此，必须去除多余的蛋白质。然而，目前用于此目的的分离技术存在决定性的缺点，例如囊泡受损、处理时间长或可处理的样品体积小。为了克服这些缺点，我们研究了Capto™ Core 700树脂用于聚合物囊泡纯化的适用性。通过中空纤维流场流分馏技术验证，聚合物囊泡通过柱子时未受到损坏或其他影响。以三种具有不同物理化学性质的蛋白质为例，结果表明，不同类型的未包裹蛋白质能从聚合物囊泡分散体中有效去除。对于所应用的蛋白质，在存在聚合物囊泡的情况下，每毫升树脂的动态结合容量在9.5至16.5毫克之间。该技术可用于小体积和大体积样品。此外，无需特殊实验室设备即可使用。这为聚合物囊泡增添了一种新的、易于使用的纯化方法，也将有助于功能化聚合物囊泡的大规模生产。
