POROUS POLYMER RODS AS CHROMATOGRAPHIC OR REACTIVE MEDIA

本提案的目标是设计、编制和测试一个 用于分离的全新形式的分离介质， 分离生物分子。 新的分离介质基于 一个多孔聚合物的连续体，直接模制到 色谱柱，而不是由小颗粒机械制成 填充到柱中。 新的分离介质具有许多优点 在易于制备、可重复性、分离通用性方面 化学，结合化学梯度的能力，紧凑性， 适用于微型诊断、制备分离， 生物反应器 连续介质的制备是简单的，相当于一个普通的 在致孔物质存在下的本体聚合，而不是 用于制备单分散的更繁琐和要求更高的技术 颗粒聚合物介质。 结果，许多不能被聚合的单体被聚合。 容易用于制备颗粒介质，包括许多 亲水性或水溶性单体特别适合 用于分离生物聚合物，可以直接使用。 的多功能性 分离化学物质的混合能力进一步增强， 在单一连续介质中的几种化学物质。 可以实现这一点 与具有互补化学性质的单体，或通过产生 柱壳内的浓度或反应性梯度 介质的制备，或者甚至通过几种介质的组合， 在单个柱内串联的化学物质。 准备的容易程度是 这是一个重要的优点，因为它消除了包装的变幻莫测， 已知其极大地影响填充柱的效率。 此外 连续介质没有特别的空隙体积 负责填充颗粒柱中的质量传递， 持续大约总柱体积的40%。 相反，连续媒体 含有包括大流通量的孔的互连阵列，或 灌注孔。 移动的相的总灌注意味着流动 是通过多孔介质本身， 而不是围绕着堆积的颗粒。 我们的目标是了解和控制 对于可重复制备连续多孔 分离介质 重点将放在效率和多功能性 应用于生物系统的分离化学 蛋白质、药物或代谢物。

The objectives of this proposal are to design, prepare and test an entirely new form of separation media for use in the separation and isolation of biological molecules. The new separation media are based on a continuous body of porous polymer molded directly into the housing of a chromatography column rather than made of small particles mechanically packed into the column. The new separation media offer many advantages in ease of preparation, reproducibility, versatility of separation chemistry, ability to incorporate gradients of chemistry, compactness, adaptability to microscale diagnostics, preparative separations, and bioreactors. The preparation of the continuous media is simple, amounting to a plain bulk polymerization in the presence of porogenic substances, rather than the more tedious and demanding techniques used to prepare monodispersed particulate polymer media. As a result, many monomers which cannot be used readily in the preparation of particulate media, including many hydrophilic or water soluble monomers that are particularly well-suited for the separation of biopolymers, can be used directly. The versatility of the separation chemistry is further enhanced by the ability to mix several chemistries in a single continuous medium. This can be achieved with monomers having complementary chemistries, or through the creation of concentration or reactivity gradients within the column housing during preparation of the media, or even by the combination of several chemistries in series within a single column. The ease of preparation is an important advantage as it removes the vagaries of packing which are known to affect greatly the efficiency of packed columns. In addition the continuous media have no interparticular void volumes which are responsible for mass transfer in packed particulate columns and account for ca. 40% of total column volume. Instead, the continuous media contain an interconnected array of pores including large flow-through, or perfusion pores. Total perfusion by the mobile phase means that the flow of the substances to be separated is through the porous medium itself, rather than around the packed particles. Our targets will be to gain an understanding and control the factors that are essential for the reproducible preparation of continuous porous separation media. Emphasis will be placed on efficiency and versatility of separation chemistry for application to biological systems such as proteins, drugs, or metabolites.