Glycoengineering insect cells for commercial recombinant glycoprotein production

用于商业重组糖蛋白生产的糖工程昆虫细胞

基本信息

批准号：
8733179
负责人：
Christoph Geisler
金额：
$ 29.68万
依托单位：
GLYCOBAC, LLC
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-30 至 2016-07-31
项目状态：
已结题

项目摘要

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.

描述：大多数人类治疗性糖蛋白（生物制剂）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，BICS 尚未用于制造非疫苗生物制剂，因为它无法生产其功效所需的人型 N-聚糖。我们之前的研究表明，我们可以通过对昆虫细胞进行糖工程改造，使其 N-糖基化途径人性化，从而解决这个问题。然而，这些学术努力并不是为了创建能够以低成本进行有效的人型 N-糖基化的最佳糖基化昆虫细胞系。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。我们将通过在原始、隔离和有据可查的条件下以新的和创新的方式扩展我们之前的努力和糖工程昆虫细胞来实现这一目标。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人类型 N-聚糖的糖蛋白。我们将在原始、分离和有据可查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，并广泛表征细胞系，以促进未来的验证和商业化工作。我们还将研究使用新的糖工程昆虫细胞系作为制造平台的可行性，通过将其生产水平与亲本细胞系进行比较，使用三种不同的糖蛋白生物制剂，其天然N-聚糖结构与通过设计生产的那些相同。或更多新的昆虫细胞系。最后，我们将通过表达和纯化每种生物并定量分析其 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 拥有保护糖基工程 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、展示其能力并在 II 期验证其安全性和有效性后，GlycoBac 将再许可这些细胞系与多家生物技术公司和合同制造组织合作。 GlycoBac 还将为全球基础和应用生物医学研究界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正确和/或可预测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。