Monolithic Media and Technology for Bioprocessing

描述(申请人提供)：这项工作的目标是为大规模生产重组蛋白开发新的制造技术。单抗等生物大分子的纯化技术是生物学研究和治疗蛋白生产的重要组成部分，这些治疗蛋白被用于越来越多的癌症治疗和其他医学应用。用于这些应用的传统吸着剂是具有大孔和大颗粒尺寸的珠状形式。需要大孔来容纳蛋白质；颗粒尺寸需要较大，以最大限度地减少缓冲流产生的压缩压降。较大的颗粒尺寸和较小的生物大分子在颗粒孔内的扩散系数降低了分离过程的效率。由于大孔尺寸的原因，这些材料是易碎和压缩的，使得它们在制备层析中很难使用。在制备规模上，为了最大限度地减少压降及其后果，这种类型的离子交换柱通常是大直径的浅床，很难包装和维护。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Macro-IPN基板大规模应用的新技术上，这些基板是由PI在NIH相关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式、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，并将使这项技术能够利用已经为大规模药物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)设计和制造基于Macro-IPN底物的离子交换设备，(2)开发基于Macro-IPN底物的材料、设备和方法，从而实现线性可扩展、更大规模的应用(3)从容量、动力学、扩散能力以及与现有技术的比较等方面表征应用于免疫球蛋白和较小蛋白质的性能。这项技术的广泛、长期目标是实现(A)一次性使用或一次性使用的治疗性蛋白质的层析系统，以及(B)将改进和简化生物处理设施中单一或多个产品的制造的系统。避免可能的交叉污染，以及在清洁协议后验证系统的需要，有望提高对良好制造工艺(GMP)标准的遵从性，并加快FDA的审批过程。潜在的商业产品是(A)阴离子和阳离子交换层析设备，(B)亲和层析设备，(C)用于大规模捕获和释放层析的低成本一次性设备，以及(D)涉及生物处理应用的膜产品的公司的特定应用许可证。 公共卫生相关性：拟议的研究与公共卫生的相关性是为加速和改进正在开发的治疗药物的制造方法提供新技术，这些药物正在开发以消除癌症造成的痛苦和死亡。这个项目的技术需要确保交付，并降低大量需要的新的抗癌治疗药物的成本，如重组蛋白

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