GOALI: Collaborative Research: Industrial Implementation of Smart Biopolymers for Purification of Biological Products

卡兰德，Pankaj/Chen，Wilfred 1403697/1403724 Rensselaer理工学院/特拉华大学拟议的项目是工业(百时美施贵宝)和学术界(Rensselaer理工学院和特拉华大学)之间的合作，旨在开发下一代最先进的智能生物聚合物系统，用于工业生物加工中蛋白质疗法的纯化。与传统的层析过程相比，在规模、成本和易于部署方面具有优势的生物材料回收新技术的开发在工业上有很大的需求和兴趣。PI建议开发结合了亲和层析的选择性以及沉淀的可扩展性和成本效益的亲和沉淀过程。该项目将采用新型亲和肽与高效的智能生物聚合物相结合，这种聚合物可以在盐和/或温度发生微小变化的情况下诱导进入和离开溶液。生物制品的非层析亲和力回收方法的发展引起了人们的极大兴趣。亲和沉淀结合了亲和层析的选择性以及沉淀的可扩展性和成本效益。在NSF支持的先前的研究中，PI使用了融合到mAb结合Z结构域(ELP-Z)的弹性蛋白样多肽来开发一种可扩展的mAb亲和沉淀方法。这项拟议的工作建立在先前工作的基础上，旨在生成全新类型的智能生物聚合物亲和试剂，并评估它们在工业上从包涵体和聚乙二醇化过程中提纯单抗、融合和非单抗生物制品的有效性。将与百时美施贵宝(BMS)合作，评估它们在大规模蛋白质纯化中的实用价值。这项研究将为生物制品的大规模纯化提供一个灵活的平台。拟议项目的第一部分将是使用ZELP-E2纳米笼在较温和的操作条件下亲和沉淀mAbs和Fc融合蛋白的新方法。该项目的第二部分将为我们的工业合作伙伴BMS提供的两类非mAb蛋白开发新型多肽亲和配体。然后，亲和多肽将被用于亲和沉淀形式，以选择性地从具有挑战性的原料中捕获产物，例如折叠蛋白质池和聚乙二醇化后反应混合物。这项工作汇集了在蛋白质工程、多肽亲和设计、亲和沉淀和下游生物处理方面的不同专业知识的Co-PI，开发了全新类别的亲和沉淀试剂(Z-ELP-E2纳米笼、ELP亲和多肽和多价亲和捕获E2-(xAPy-ELP))，如果成功，可能会产生一个新的集成平台，大大简化生物制品的回收和纯化。这项拟议的研究可能会对以单抗、融合蛋白、包涵体生物制品和聚乙二醇化蛋白为代表的广泛治疗产品的亲和力捕获产生巨大影响。这些新技术的结合可能代表着生物制品下游加工的全新方法，具有重大的长期工业影响。这项工作的成功不仅可能影响难以回收和合成修饰的生物制品的加工，而且还可能为许多正在开发的许多新生物制品的生物加工建立新的范例。学术界和产业界之间的这种合作将作为一项概念验证研究，以期在工业上实现这一用于生物产品纯化的变革性技术。讨论了通过将研究生、本科生和K12纳入研究机会而对教育产生影响的机制。这一提出的分子水平和宏观分离水平的培训受到生物技术行业的高度追捧。

Karande, Pankaj / Chen, Wilfred 1403697 / 1403724 Rensselaer Polytechnic Institute / University of Delaware The proposed project is a collaboration between industry (Bristol-Myers Squibb) and academia (Rensselaer Polytechnic Institute and University of Delaware) aimed at developing the next generation and state-of-art smart biopolymers system for the purification of protein therapeutics in industrial bioprocessing. There is a significant need and interest in the industry for the development of novel techniques for recovery of biological materials that are advantageous in terms of scale, cost, and ease-of-deployment compared to conventional chromatographic processes. The PIs propose to develop affinity precipitation processes that combine the selectivity of affinity chromatography along with the scalability and cost benefits of precipitation. This project will employ new classes of affinity peptides in concert with efficient smart biopolymers which can be induced to come in and out of solution with minor changes in salt and/or temperature. There is significant interest in the development of non-chromatographic affinity recovery processes for biological products. Affinity precipitation combines the selectivity of affinity chromatography along with the scalability and cost benefits of precipitation. In prior research supported by NSF the PIs have employed Elastin like polypeptides fused to the mAb binding Z domain (ELP-Z) to develop a scalable mAb affinity precipitation method. The proposed work builds upon the previous work to generate entirely new classes of smart biopolymer affinity reagents and evaluate their utility for the industrial purification of mAbs, FCfusions, and non-mAb biologics from inclusion body and pegylation processes. Their practical utility will be evaluated for large-scale protein purification in partnership with Bristol-Myers Squibb (BMS). This research will result in a flexible platform for large-scale purification of biological products. The first component of the proposed project will be a new approach for affinity precipitation of mAbs and Fc fusion proteins under milder operating conditions using ZELP-E2 nanocages. The second component of the project will develop novel peptide affinity ligands for two classes of non-mAb proteins provided by our industrial collaborator BMS. The affinity peptides will then be employed in affinity precipitation formats to selectively capture the products from challenging feed stocks such as refolded protein pools and post-pegylation reaction mixtures. This work brings together Co-PIs with diverse expertise in protein engineering, peptide affinity design, affinity precipitation and downstream bioprocessing to develop entirely new classes of affinity precipitation reagents (Z-ELP-E2 nanocages, ELPaffinity peptides, and polyvalent affinity capture E2-(xAPy-ELP)) which if successful may result in a new integrated platform that will greatly simplify the recovery and purification of biological products. The proposed research could have a dramatic impact on the affinity capture of a broad range of therapeutic products represented by monoclonal antibodies, fusion proteins, biologics from inclusion bodies, and pegylated proteins. The combination of these novel technologies could represent entirely new approaches for the downstream processing of biologics with significant long-term industrial impact. Success in this work could not only impact the processing of difficult to recover and synthetically modified biologics but may also establish new paradigms for the bioprocessing of a wide range of many new biologics in development. This collaboration between academia and industry will serve as a proof-of-concept study for the potential industrial implementation of this transformative technology for the purification of biological products. Mechanisms for impact on education via graduate, undergraduate and K12 inclusion in research opportunities are discussed. This proposed molecular level and macroscopic separations level training is highly sought after by the biotechnology industry.