GlycoFlow Glycoengineering of therapeutic antibodies by flow biocatalysis.

GlycoFlow 通过流动生物催化对治疗性抗体进行糖工程。

• 批准号: 571434-2021
• 负责人: Cecioni, Samy
• 金额: $ 3.28万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=729162
• 关键词: GlycoFlow; Glycoengineering; therapeutic; antibodies; flow

The advent of biopharmaceuticals is revolutionizing health care, opening up entirely new avenues against diseases that are difficult to treat using traditional drugs. Marketed therapeutic antibodies are proteins that recognize and bind specific molecules on diseased cells, thereby eliciting a beneficial immunological response. A lesser-known fact is that antibodies are usually not single homogenous proteins. They also require glycosylation, which means the antibody is linked to a complex structure of various sugars (i.e. glycans). This glycosylation is essential for generating efficacious therapeutic antibodies. Therefore, these biopharmaceuticals are manufactured using elaborate and costly protein expression systems that carry the biological machinery needed to install these sugars. Because living cells are used, the resulting product comprises a heterogenous population with multiple glycan structures that can show significant differences, with up to a 50-fold decrease in efficacy when the glycans are fucosylated (decorated with a fucose sugar). Such a heterogeneity imposes a significant burden on the development and use of therapeutic antibodies, insofar as it impacts efficacy and safety profiles, manufacturing costs, batch variability as well as regulatory requirements.Consequently, better approaches are needed to control the structures of glycans linked to antibodies, as well as the consistency of the glycan population. Current strategies exploit genetically modified expression cell lines (e.g. to reduce the level of fucosylation) that still produce highly variable glycosylation profiles. Alternatively, in vitro batch chemo-enzymatic modifications have proven successful at generating homogenously glycosylated antibodies, but they suffer from complex purification processes and scalability issues.We propose a GlycoFlow platform in which therapeutic antibodies could be glycoengineered using flow biocatalysis. By passing a solution of antibodies through modular cartridges of glycan-processing enzymes, we aim to manipulate the structures of sugars directly in continuous flow, generating optimized and homogenous glycosylated antibodies. Continuous flow technologies are game-changers for the modular synthesis of pharmaceutical ingredients with high efficiency and consistency, but have yet to be leveraged for glycoengineering of biopharmaceuticals.To achieve this goal, we have assembled a diverse team of young and experienced experts in glycosciences, glycan-processing enzymes, antibody biochemistry, and protein engineering. Our team will take advantage of the historical expertise in flow chemistry at UdeM, coupled with world-leading researchers from Concordia and McMaster to deliver innovative solutions for the next-generation of biopharmaceuticals. Specifically, we will develop flow technologies that yield homogenously glycosylated antibodies as well as simpler solutions towards afucosylated and sialylated glycans, both of which have showed increased efficacy and improved anti-inflammatory properties.

生物制药的出现正在彻底改变医疗保健，为传统药物难以治疗的疾病开辟了全新的途径。市售的治疗性抗体是识别并结合患病细胞上的特异性分子的蛋白质，从而引发有益的免疫应答。一个鲜为人知的事实是，抗体通常不是单一的同质蛋白质。它们还需要糖基化，这意味着抗体与各种糖（即聚糖）的复杂结构连接。这种糖基化对于产生有效的治疗性抗体是必不可少的。因此，这些生物药物是使用精心制作和昂贵的蛋白质表达系统制造的，该系统携带安装这些糖所需的生物机器。由于使用活细胞，所得产物包含具有多种聚糖结构的异源群体，这些聚糖结构可显示出显著差异，当聚糖被岩藻糖基化（用岩藻糖修饰）时，功效降低高达50倍。这种异质性给治疗性抗体的开发和使用带来了巨大的负担，因为它会影响疗效和安全性、生产成本、批次可变性以及监管要求。因此，需要更好的方法来控制与抗体连接的聚糖的结构以及聚糖群体的一致性。目前的策略利用遗传修饰的表达细胞系（例如，以降低岩藻糖基化水平），其仍然产生高度可变的糖基化谱。另外，在体外批量化学-酶修饰已被证明成功地产生同质糖基化抗体，但他们遭受复杂的纯化过程和scalability issues.We提出了一个GlycoFlow平台，其中治疗性抗体可以使用流动生物催化糖工程。通过使抗体溶液通过聚糖加工酶的模块化柱，我们的目标是在连续流中直接操纵糖的结构，产生优化的和同质的糖基化抗体。连续流技术是高效、稳定的药物成分模块化合成的革命性技术，但在生物制药的糖工程中尚未得到充分利用。为了实现这一目标，我们组建了一支由糖科学、聚糖加工酶、抗体生物化学和蛋白质工程领域的年轻而经验丰富的专家组成的多元化团队。我们的团队将利用UdeM在流动化学方面的历史专业知识，以及来自Concordia和McMaster的世界领先研究人员，为下一代生物制药提供创新解决方案。具体来说，我们将开发流技术，产生均匀糖基化的抗体以及更简单的解决方案，对无岩藻糖基化和唾液酸化聚糖，这两者都显示出增加的功效和改善的抗炎特性。