Monolithic Media and Technology for Bioprocessing

描述（由申请人提供）：本工作的目的是开发用于大规模生产重组蛋白的新生产技术。大生物分子（如单克隆抗体）的纯化技术是生物学研究的重要组成部分，也是生产治疗性蛋白质的重要组成部分，这些蛋白质用于越来越多的癌症治疗和其他医学应用。 用于这些应用的常规吸附剂为珠粒形式，其特征在于具有大孔和大粒度。需要大孔来容纳蛋白质;颗粒尺寸需要大以最小化缓冲液流动产生的压缩压降。大颗粒尺寸和颗粒孔内大生物分子的小扩散系数降低了分离过程的效率。由于大的孔径，这些材料是易碎的和可压缩的，使得它们难以用于制备色谱。在制备规模上，为了使压降及其后果最小化，具有这种类型介质的离子交换柱通常是难以填充和维护的大直径浅床。这项研究的创新将解决现有技术中的这些固有问题。 这些研究将集中在大规模应用Macro-IPN基质的新技术上，Macro-IPN基质是由PI在相关NIH项目中开发的由高容量水凝胶和惰性非织造材料组成的整体，大孔，片状材料。作为该项目的一部分开发的新系统和设备将使Macro-IPN介质的大规模应用成为可能，并将使该技术能够利用已经为大规模制药过滤器产品和应用建立的知识体系。该项目的具体目标是：（1）设计和构建基于Macro-IPN基材的离子交换装置;（2）开发基于Macro-IPN基材的材料、设备和方法，以实现线性可扩展的更大规模应用;（3）表征免疫球蛋白和较小蛋白质在容量、动力学、扩散率方面的应用性能，并与现有技术进行比较。该技术的广泛的长期目标是实现（a）用于治疗性蛋白质的一次性或一次性色谱系统和（B）将改进和简化生物加工设施中的单一或多种产品的制造的系统。避免可能的交叉污染和需要在清洁协议后验证系统，有望提高对良好生产工艺（GMP）标准的合规性，并加快FDA的批准过程。潜在的商业产品有：（a）阴离子和阳离子交换色谱装置，（B）亲和色谱装置，（c）低成本、一次性大规模捕获和释放色谱装置，以及（d）向涉及生物处理应用的膜产品的公司颁发特定应用许可证。 公共卫生相关性：拟议的研究与公共卫生的相关性是提供新技术，加速和改进正在开发的治疗药物的制造方法，以消除癌症造成的痛苦和死亡。需要该项目的技术来确保交付并降低新的抗癌治疗方法的成本，例如大量需要的重组蛋白质

DESCRIPTION (provided by applicant): The objective of this work is to develop new manufacturing technology for large-scale production of recombinant proteins. Purification technology for large biomolecules such as monoclonal antibodies is an important component of biological research and for the production of therapeutic proteins that are used in a growing number of cancer therapies and other applications in medicine. Conventional sorbents for these applications are in the form of beads which characteristically have large pores and a large particle size. The large pores are needed to accommodate the proteins; the particle size needs to be large to minimize the compressive pressure drops produced by buffer flow. The large particle size and small diffusion coefficients of large biomolecules within the particle pores reduces efficiency of the separation process. As a result of the large pore size, these materials are fragile and compressive making their use in preparative chromatography difficult. On a preparative scale, in order to minimize pressure drop and its consequences, ion exchange columns with this type of media are typically large diameter, shallow beds that are difficult to pack and maintain. Innovations from this research will address these inherent problems in the existing technology. These studies will focus on new technology for large scale application of Macro-IPN substrates which are monolithic, macroporous, sheet-form materials consisting of a high capacity hydrogel and an inert nonwoven material which were developed by the PI in a related NIH project. New systems and devices developed as part of this project will enable large scale applications of the Macro-IPN media and will allow the technology to capitalize on the body of knowledge that has already been established for large-scale pharmaceutical filter products and applications. The specific aims of this project are, (1) Design and build ion exchange devices based on Macro-IPN substrates, (2) Develop materials, equipment, and methods based on Macro-IPN substrates that will lead to linearly scalable, larger scale applications (3) Characterize performance for applications to immunoglobulins and smaller proteins in terms of capacities, kinetics, diffusivities, and comparisons to existing technology. The broad, long term objectives for this technology are to enable (a) one- time-use or disposable chromatographic systems for therapeutic proteins and (b) systems that will improve and simplify the manufacture of single or multiple products in a bio-processing facility. The avoidance of possible cross contamination and the need to validate systems after cleaning protocols promises to improve compliance with Good Manufacturing Process (GMP) standards and speed up the FDA approval process. Potential commercial products are (a) devices for anion and cation exchange chromatography, (b) devices for affinity chromatography, (c) low cost, disposable devices for large scale capture and release chromatography, and (d) application specific licenses to companies involved in membrane products for bio-processing applications. PUBLIC HEALTH RELEVANCE: The relevance of the proposed research to public health is to provide new technology for accelerated and improved manufacturing methods for therapeutic drugs that are being developed to eliminate suffering and death due to cancer. The technology from this project is needed to ensure delivery and reduce the cost of new, anti-cancer therapeutics such as recombinant proteins which are needed in large quantities