Coiled-coil assisted control of antibody quality, delivery and binding

治疗性重组蛋白的市场现在由单克隆抗体（mAb）主导。随着超过60种mAb获得批准，超过350种正在进行临床试验，这个市场现在超过500亿美元。由于mAb生产模式的复杂性，mAb生产仍然是公共卫生保健系统的重大经济负担。 一方面，在生物制药行业，缺乏成本效益和高通量工程解决方案来评估mAb关键质量属性，严重阻碍了依赖细胞培养的当前生产工艺的改进。首先，mAb糖基化（由细胞进行的翻译后修饰）的评估至关重要，因为糖基化直接影响mAb稳定性和治疗功效。 另一方面，在临床中，通过静脉途径施用mAb的模式也需要改进（更可能通过材料工程）以实现最大治疗功效。实际上，静脉内施用迫使使用大剂量，进而导致患者不便和高医疗成本。缺乏有效和通用的方法来促进mAb单独或与其他药物组合的持续局部递送，严重阻碍了下一代治疗剂的开发和实施。 我正在进行的研究计划的重点是开发通用和有效的工程策略，以控制重组蛋白质的质量和交付。在过去的几年里，我的团队在解决这些问题方面取得了重大进展：我们率先使用了依赖于两种不同肽的移植策略，即，E和K线圈，以设计更好的生物传感测定和控制生物活性蛋白质捕获到水凝胶中。由于单克隆抗体的重要性，我们将在未来5年内专门关注这类生物分子。我们对这项发现补助金的短期目标是： 1)开发生物传感和数值工具来表征mAb糖基化，从而为mAb生物制造创建增强的过程分析技术 2)设计可调的自组装水凝胶，用于控制mAb和其他实体的释放，从而满足行业对提供联合疗法平台的需求。 3)通过微调E和K螺旋肽序列，进一步完善我们的移植和递送技术平台，扩展其相互作用的表征，以支持该计划的短期和长期目标。 总的来说，这个发现补助金计划将有助于培养10名本科生和4名博士生。实现我们的研究目标不仅将深入了解mAb糖基化和控释的基本原理，还将促进技术转让给我们的加拿大工业合作伙伴，最终为加拿大人提供更有效和更具成本效益的治疗。

The market of therapeutic recombinant proteins is now dominated by monoclonal antibodies (mAbs). With more than 60 mAbs approved and more than 350 undergoing clinical trials, this market now exceeds 50 billion dollars. Due to the complexity of mAb mode of production, mAb manufacturing still represent a significant economic burden to the public health care system. On the one hand, in the biopharmaceutical industry, the lack of cost-effective and high-throughput engineering solutions to assess mAb critical quality attributes severely impedes the improvement of current manufacturing processes relying on cell culture. First and foremost, the assessment of mAb glycosylation, a post translational modification performed by the cells, is crucial as glycosylation directly influences mAb stability and therapeutic efficacy. On the other hand, in the clinic, the mode of administration of mAbs via intravenous route, also needs improvement (more likely via materials engineering) to achieve maximal therapeutic efficacy. Indeed, intravenous administration forces the use of huge doses, in turn leading to patient inconvenience and high medical treatment costs. The lack of an efficient and versatile approach to promote the sustained local delivery of mAbs, alone or in combination with other drugs, severely impedes the development and implementation of the next-generation therapeutics. The focus of my ongoing research program is to develop versatile and efficient engineering strategies to control recombinant protein quality and delivery. Over the past years, my team has made significant progress on addressing these issues: we spearheaded the use of a grafting strategy relying on two distinct peptides, i.e., the E and K coils, to design better biosensing assays and control the capture of bioactive proteins into hydrogels. Because of the importance of mAbs, we will focus the next 5 years specifically on this class of biomolecules. Our short-term objectives for this Discovery Grant are to: 1) Develop the biosensing and numerical tools to characterize mAb glycosylation, and thus create an enhanced process analytical technology for mAb biomanufacturing 2) Design tunable, self-assembled hydrogels for the controlled release of mAbs and other entities, hence addressing the needs of the industry for platforms delivering combined therapies. 3) Further refine our grafting and delivery technological platform by fine-tuning the E and K coil peptide sequences, extending the characterization of their interactions to support the short and long-term objectives of this program. Overall, this Discovery Grant Program will contribute to the training of 10 undergraduate and 4 PhD students. Attaining our research objectives will not only yield insight into the fundamentals of mAb glycosylation and controlled release but facilitate technology transfer to our Canadian industrial partners, ultimately leading to more efficient and cost- effective treatment to Canadians.