Glycoengineering insect cells for commercial recombinant glycoprotein production

描述：大多数人类治疗性糖蛋白(生物制品)是使用哺乳动物细胞系制造的，这些细胞系可以添加临床疗效所需的人型多糖。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，它们可以含有不定物，它们可以产生免疫原性多糖。杆状病毒-昆虫细胞系统(BICS)提供了一个有吸引力的替代方案，因为它快速、安全，并且已经被用于生产两种FDA许可的疫苗。尽管有这些优点，BICS还没有被用于生产非疫苗生物制品，因为它不能生产其疗效所需的人型N-聚糖。我们之前的研究表明，我们可以通过糖工程来人源化昆虫细胞的N-糖基化途径来解决这个问题。然而，这些学术努力并不是为了创造能够以低成本高效地进行人型N-糖基化的最佳糖工程昆虫细胞系。因此，这个实验室到市场项目的总体目标是创造新的糖工程昆虫细胞系，能够提供高效的人型N-糖基化，而不需要昂贵的介质补充剂，用于商业生物制品制造和基础生物医学研究。我们将通过扩展我们之前的努力，在原始、隔离和有充分记录的条件下，以新的和创新的方式改造昆虫细胞，从而实现这一目标。更具体地说，我们将分离三个新的昆虫细胞系，这些细胞系旨在生产具有商业生物制品中常见的独特的人型N-糖聚糖的糖蛋白。我们将在原始、隔离和有良好记录的条件下生产和维护这些细胞系。然后，我们将生产细胞库，测试它们的污染物，并广泛地表征细胞系，以促进未来的验证和商业化努力。我们还将研究将新的糖工程昆虫细胞系用作制造平台的可行性，方法是将它们的生产水平与使用三种不同糖蛋白生物制品的亲本细胞系进行比较，这些糖蛋白生物制品的天然N-糖链结构与一个或多个新昆虫细胞系设计生产的相同。最后，我们将通过表达和纯化每种生物并定量描述它们的N-糖链结构，来检验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BICS关键知识产权的独家许可。在创建了这里描述的新细胞系，展示了它们的能力，并在第二阶段验证了它们的安全性和有效性后，GlycoBac将对这些细胞系进行再许可 与多家生物技术公司和代工组织建立联系。GlycoBac还将为世界各地的基础和应用生物医学研究社区提供对这些新细胞系的开放访问。因此，该项目产生的工具将对人类医学和需要适当和/或可预测的糖基化重组糖蛋白的基础生物医学研究的不同领域产生重大和广泛的影响。

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.