Novel Chromatographic Sorbents for Large Biomolecules

描述(申请人提供)：大生物分子的纯化是生物学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许多应用。用于此目的的常规层析介质具有大(大于5纳米)的孔来容纳这些大分子。因此，这些介质非常脆弱和可压缩，使得它们在柱层析和生物处理中的使用非常困难。此外，为了将缓冲流产生的压缩压降降到最小，介质的颗粒尺寸需要很大，通常直径超过100微米。大分子在颗粒内部的大尺寸和极小的扩散系数极大地降低了分离过程的效率。该项目的目标是开发一种新型的色谱介质，这种介质可以通过在非织造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。所得到的层析介质将具有高效率和坚固性的理想组合，使其用作层析介质比用于类似应用的现有介质更有效、更高效、成本更低。具体目标是(1)开发具有离子交换功能、高容量和高传质速率的聚脲涂层的非织造布系列，用于重组蛋白质、单抗和较小的蛋白质的高传质；(2)利用流动设备表征涂层非织造布对生物分子的容量、动力学和扩散性；以及(3)基于涂层非织造布的特性开发用于色谱装置的设计和制造原型。

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.