Novel Chromatographic Sorbents for Large Biomolecules

描述（由申请人提供）：大生物分子的纯化是生物研究和重组蛋白生产的重要组成部分，重组蛋白在医学上有多种应用。用于此目的的传统色谱介质具有大（大于 5 纳米）的孔来容纳这些大分子。因此，这些介质极其脆弱且具有压缩性，使得它们在柱色谱和生物加工中的使用非常困难。此外，为了最小化缓冲流产生的压缩压降，介质的颗粒尺寸需要很大，通常直径超过100微米。颗粒孔内大分子的大尺寸和极小的扩散系数，大大降低了分离过程的效率。该项目的目标是开发一种新型色谱介质，该介质可以通过在无纺布上涂覆非常薄的聚合物涂层（小于6微米）并具有适合吸附大生物分子的离子交换基团来制备。所得的色谱介质将具有所需的高效率和稳健性的组合，使得它们作为色谱介质的使用比用于类似应用的现有介质更有效、更高效且成本更低。具体目标是（1）开发一系列具有聚脲涂层的无纺布，具有离子交换功能、高容量和重组蛋白、单克隆抗体和较小蛋白质的高传质率，（2）使用流过装置表征涂层无纺布的容量、动力学和生物分子扩散性，以及（3）根据涂层无纺布的特性开发色谱装置的设计和原型。

DESCRIPTION (provided by applicant): The purification of large biomolecules is an important component of biological research and for the production of recombinant proteins that are being utilized for numerous applications in medicine. The conventional chromatographic media used for this purpose have large (greater than 5 nanometer) pores to accommodate these large molecules. As a consequence these media are extremely fragile and compressive, making their use in column chromatography and bioprocessing very difficult. Furthermore, in order to minimize the compressive pressure drops produced by buffer flow, the particle size of the media need to be large, typically more than 100 micrometers in diameter. The large size and the extremely small diffusion coefficients of large molecules within the pores of the particles, greatly reduces the efficiency of the separation processes. The goal of this project is focused on the development of a new type of chromatographic medium which can be prepared by applying very thin polymeric coatings (less than 6 micrometers) onto non-woven fabrics and having ion exchange groups that are suitable for the adsorption of large biomolecules. The resulting chromatographic media will have the desired combination of high efficiency and robustness, making their use as chromatographic media more effective, more efficient, and at lower cost than existing media for similar applications. The specific aims are (1) develop a family of non-wovens with polyurea coatings having ion exchange functionalities, high capacities, and high rates of mass transfer for recombinant proteins, monoclonal antibodies, and smaller proteins, (2) characterize the coated non-wovens in terms of their capacities, kinetics, and diffusivities for biomolecules using flow through devices, and (3) develop designs and build prototypes for chromatographic devices based on the properties of the coated non-wovens.