SBIR Phase I: Reducing the cost of biomanufacturing and biopharmaceutical production with smart biocatalysts that can be incorporated into continuous processing technologies

这个小企业创新研究（SBIR）第一阶段项目的更广泛的影响/商业潜力将是开发一种酶固定化平台，用于生物制药和其他生物制造工艺的生产，以减少与发现和制造生物治疗药物相关的时间和成本。近年来，生物制药的销售额飙升，医疗保健当局加大了对制药商的压力，要求他们降低生物制剂的成本，这使得制药工程师们专注于降低生物生产成本。为了实现这一目标，制药公司正在积极研究连续加工技术，以减少与生物制药生产相关的资本和其他成本。本提案中描述的固定化平台将提供一种与连续处理策略兼容的技术，以减少与发现和制造生物治疗药物相关的时间和成本。此外，该平台还具有降低成本和提高其他几个行业生物制造过程效率的潜力，包括食品、皮革和纺织品。这个SBIR一期项目的智力优势是开发用于生物制药生产的pH响应固定化酶。与该项目相关的三个关键技术挑战：固定化酶必须表现出与生物制剂生产相容的pH响应行为（pH 6-7）；蛋白质底物的传质必须与天然酶相当；并且固定化酶要长期保存，需要延长其货架期。为了缓解这些挑战，该项目旨在将酶与pH响应聚合物共价结合，该聚合物的配制具有接近中性pH的可逆溶解度。固定化生物催化剂的溶解度可以严格控制在特定的pH范围内，从而允许生物治疗产品的连续处理和易于纯化。传质不良往往是由于固定化生物催化剂在固体载体上造成的。这些挑战在生物偶联反应中被进一步夸大，因为偶联伙伴蛋白质通常比小分子更容易受到立体结构的影响。因此，固体载体的制造将被探索，以生产不同尺寸、形态和溶解性质的树脂，这些树脂与大分子功能化相兼容。此外，本项目生成的固定化酶可以在可溶性条件下利用，结合了固定化和可溶性酶催化的积极属性。开发商业上可行的催化剂保质期的最后挑战将通过开发新的干燥方案来解决。将开发货架稳定的固定化酶，使库存可以保持，而不需要长时间的催化剂生产。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project will be to develop an enzyme immobilization platform to be used in the production of biopharmaceuticals and other biomanufacturing processes to reduce the timeline and cost associated with discovering and manufacturing biotherapeutics. Biopharmaceutical sales have soared in recent years and healthcare authorities have increased pressure on pharmaceutical manufacturers to reduce the cost of biologics leaving pharmaceutical engineers to focus on reducing bioproduction costs. To accomplish this goal, pharmaceutical companies are aggressively investigating continuous processing technologies to reduce the capital and other costs associated with biopharmaceutical manufacture. The immobilization platform described in this proposal will provide a technology that is compatible with continuous processing strategies to reduce the timeline and cost associated with discovering and manufacturing biotherapeutics. In addition, this platform has the potential to reduce the cost and increase the efficiency of biomanufacturing processes across several other industries including food, leather, and textiles. The intellectual merit of this SBIR Phase I project is to develop pH responsive immobilized enzymes for the production of biopharmaceuticals. There are three key technical challenges associated with this project: The immobilized-enzyme must demonstrate pH responsive behavior compatible with biologics manufacture (pH 6-7); mass transfer of protein substrates must be comparable to the native enzyme; and extended shelf-life of the immobilized-enzyme is required for long-term storage. To mitigate these challenges, this project aims to covalently bind an enzyme to a pH responsive polymer that is formulated to have reversible solubility near neutral pH. The solubility of the immobilized-biocatalyst can be tightly controlled within a specified pH range allowing for continuous processing and facile purification of the biotherapeutic product. Poor mass transfer often results from immobilizing biocatalysts on solid supports. These challenges are further exaggerated in bioconjugation reactions because the coupling partner, a protein, is generally more affected by sterics than small molecules. Therefore, fabrication of the solid support will be explored to produce resins of varied size, morphologies, and dissolution properties that are compatible with macromolecule functionalization. Additionally, the immobilized enzyme generated in this project can be utilized under soluble conditions, which combines the positive attributes of both immobilization and soluble enzyme catalysis. The final challenge of developing a commercially viable shelf-life of the catalyst will be addressed through development of novel drying protocols. Shelf-stable immobilized enzymes will be developed so that inventory can be maintained without requiring long lead times for catalyst production.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.