Chenette Heather C S, Husson Scott M
Department of Chemical and Biomolecular Engineering and Center for Advanced Engineering Fibers and Films, Clemson University, Clemson, SC 29634, USA.
J Appl Polym Sci. 2015 Jun 5;132(21):1-7. doi: 10.1002/app.41437.
This work details the design and testing of affinity membrane adsorbers for lectin purifications that incorporate glucose-containing glycopolymers. It is the selective interaction between the sugar residues of the glycopolymer and the complementary carbohydrate-binding domain of the lectin that provides the basis for the isolation and purification of lectins from complex biological media. The design approach used in these studies was to graft glycopolymer 'tentacles' from macroporous regenerated cellulose membranes by atom transfer radical polymerization. As shown in earlier studies, this design approach can be used to prepare high-productivity membrane adsorbers. The model lectin, concanavalin A (conA), was used to evaluate membrane performance in bind-and-elute purification, using a low molecular weight sugar for elution. The membrane capacity for binding conA was measured at equilibrium and under dynamic conditions using flow rates of 0.1 and 1.0 mL/min. The first Damkohler number was estimated to relate the adsorption rate to the convective mass transport rate through the membrane bed. It was used to assess whether adsorption kinetics or mass transport contributed the primary limitation to conA binding. Analyses indicate that this system is not limited by the accessibility of the binding sites, but by the inherent rate of adsorption of conA onto the glycopolymer.
这项工作详细介绍了用于凝集素纯化的亲和膜吸附剂的设计和测试,该吸附剂包含含葡萄糖的糖聚合物。正是糖聚合物的糖残基与凝集素的互补碳水化合物结合结构域之间的选择性相互作用,为从复杂生物介质中分离和纯化凝集素提供了基础。这些研究中使用的设计方法是通过原子转移自由基聚合从大孔再生纤维素膜上接枝糖聚合物“触手”。如早期研究所示,这种设计方法可用于制备高生产率的膜吸附剂。使用模型凝集素伴刀豆球蛋白A(ConA),以低分子量糖进行洗脱,来评估膜在结合-洗脱纯化中的性能。在平衡状态下以及使用0.1和1.0 mL/min的流速在动态条件下测量膜结合ConA的容量。估算了第一个达姆科勒数,以关联吸附速率与通过膜床的对流传质速率。它用于评估吸附动力学或传质是否是ConA结合的主要限制因素。分析表明,该系统不受结合位点可及性的限制,而是受ConA在糖聚合物上的固有吸附速率的限制。