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

这个小型企业创新研究(SBIR)第一阶段项目的更广泛的影响/商业潜力将是开发一个酶固定化平台，用于生产生物制药和其他生物制造工艺，以减少与发现和制造生物疗法相关的时间和成本。近年来，生物制药销量飙升，医疗当局加大了对制药商的压力，要求它们降低生物制品的成本，这让制药工程师专注于降低生物制品的成本。为了实现这一目标，制药公司正在积极研究连续加工技术，以降低与生物制药制造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疗法相关的时间和成本。此外，该平台有可能降低食品、皮革和纺织等其他几个行业的生物制造过程的成本和效率。这个SBIR第一阶段项目的智力优势是开发pH响应型固定化酶用于生物制药的生产。这个项目有三个关键的技术挑战：固定化酶必须表现出与生物制品生产(pH 6-7)兼容的pH响应行为；蛋白质底物的传质必须与天然酶相当；为了长期储存，固定化酶必须延长货架期。为了缓解这些挑战，该项目旨在将酶与pH响应性聚合物共价结合，该聚合物的配方在中性pH附近具有可逆的溶解性。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加工和简单纯化。生物催化剂固定在固体载体上往往会导致传质效果不佳。这些挑战在生物偶联反应中被进一步夸大，因为偶联伙伴，一种蛋白质，通常比小分子更受空间结构的影响。因此，将探索固体载体的制备，以生产与高分子官能化兼容的不同尺寸、形态和溶解性能的树脂。此外，本项目产生的固定化酶可以在可溶条件下使用，这结合了固定化和可溶酶催化的优点。开发商业可行的催化剂货架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，以便在不需要长时间生产催化剂的情况下维护库存。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.