Glycoengineering insect cells for commercial recombinant glycoprotein production

产品说明：大多数人类治疗性糖蛋白（生物制品）都是使用哺乳动物细胞系制造的，这些细胞系可以添加其临床功效所需的人型聚糖。然而，哺乳动物细胞有一些严重的缺点-建立生产线需要很长时间，它们可能含有外源因子，并且它们可以产生免疫原性聚糖。杆状病毒-昆虫细胞系统（BICS）提供了一种有吸引力的替代方案，因为它快速，安全，并且已经用于生产两种FDA许可的疫苗。尽管有这些优点，但BICS尚未用于生产非疫苗生物制剂，因为它不能产生其功效所需的人型N-聚糖。我们以前的研究表明，我们可以通过糖工程昆虫细胞来解决这个问题，使其N-糖基化途径人源化。然而，这些学术努力并没有被设计成以低成本创建能够有效的人型N-糖基化的最佳糖工程化昆虫细胞系。因此，该实验室到市场项目的总体目标是创建新的糖工程昆虫细胞系，该细胞系可以提供有效的人类型N-糖基化，而无需昂贵的培养基补充剂，用于商业生物制剂制造和基础生物医学研究。我们将通过扩展我们以前的努力和糖工程昆虫细胞在原始的，孤立的和有据可查的条件下，以新的和创新的方式实现这一目标。更具体地说，我们将分离出三种新的昆虫细胞系，这些细胞系旨在生产具有商业生物制剂中常见的独特的人型N-聚糖的糖蛋白。我们将在原始、隔离和记录良好的条件下生产和维持这些细胞系。然后，我们将生产细胞库，测试它们的污染物，并广泛表征细胞系，以促进未来的验证和商业化工作。我们还将通过使用三种不同的糖蛋白生物制剂（天然N-聚糖结构与一种或多种新昆虫细胞系设计生产的N-聚糖结构相同），将其生产水平与亲本细胞系进行比较，研究使用新糖工程昆虫细胞系作为生产平台的可行性。最后，我们将通过表达和纯化每种生物制剂并定量分析其N-聚糖结构来检查我们的新昆虫细胞系的人型N-糖基化效率。GlycoBac，LLC拥有保护糖工程化BICS的关键知识产权的独家许可。在创建本文所述的新细胞系，证明其能力，并在II期中验证其安全性和有效性后，GlycoBac将再许可这些细胞系。 生产线，以多个生物技术公司和合同制造组织。GlycoBac还将为全球基础和应用生物医学研究社区提供这些新细胞系的开放获取。因此，该项目中产生的工具将对人类医学和需要适当和/或可预测糖基化重组糖蛋白的基础生物医学研究的各个领域产生重大而广泛的影响。

DESCRIPTION: Most human therapeutic glycoproteins (biologics) are manufactured using mammalian cell lines that can add the human-type glycans required for their clinical efficacy. However, mammalian cells have some serious disadvantages - it takes a long time to establish production lines, they can contain adventitious agents, and they can produce immunogenic glycans. The baculovirus-insect cell system (BICS) offers an attractive alternative because it is fast, safe, and already being used to manufacture two FDA-licensed vaccines. Despite these advantages, the BICS is not yet being used to manufacture non-vaccine biologics because it cannot produce the human-type N-glycans needed for their efficacy. Our previous research has shown that we can solve this problem by glycoengineering insect cells to humanize their N-glycosylation pathways. However, those academic efforts were not designed to create optimally glycoengineered insect cell lines capable of efficient human-type N-glycosylation at low cost. Thus, the overall goal of this Lab to Marketplace project is to create new glycoengineered insect cell lines that can provide efficient human-type N-glycosylation without costly media supplements for use in commercial biologics manufacturing and basic biomedical research. We will achieve this goal by extending our previous efforts and glycoengineering insect cells in new and innovative ways under pristine, isolated, and well-documented conditions. More specifically, we will isolate three new insect cell lines designed to produce glycoproteins with distinct, human-type N-glycans commonly found on commercial biologics. We will produce and maintain these cell lines under pristine, isolated, and well-documented conditions. We will then produce cell banks, test them for contaminants, and extensively characterize the cell lines to facilitate future validation and commercialization efforts. We also will examine the feasibility of using the new glycoengineered insect cell lines as manufacturing platforms by comparing their production levels to the parental cell line using three different glycoprotein biologics with native N-glycan structures identical to those to be produced, by design, by one or more of the new insect cell lines. Finally, we will examine the human-type N-glycosylation efficiencies of our new insect cell lines by expressing and purifying each biologic and quantitatively profiling their N-glycan structures. GlycoBac, LLC has the exclusive license to key intellectual property protecting glycoengineered BICS. After creating the new cell lines described herein, demonstrating their capabilities, and, validating their safety and efficacy in Phase II, GlycoBac will sublicense these lines to multiple biotechnology companies and contract manufacturing organizations. GlycoBac also will provide open access to these new cell lines for the worldwide basic and applied biomedical research communities. Thus, the tools generated in this project will have a significant and broad impact on human medicine and diverse areas of basic biomedical research requiring properly and/or predictably glycosylated recombinant glycoproteins.