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

• 批准号: 8589209
• 负责人: Christoph Geisler
• 金额: $ 30.24万
• 依托单位: GLYCOBAC, LLC
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8589209
• 关键词: Acute; Address; Area; Baculoviruses; Biological; Biomedical Research; Biotechnology; Carbohydrates; Cell Line; Cells; Communities; Contracts; Disadvantaged; Engineering; Future; Genetic Engineering; Glycoproteins; Goals; Human; Industry; Insecta; Intellectual Property; Letters; Licensing; Mammalian Cell; Medicine; Pathway interactions; Pharmaceutical Preparations; Phase; Polysaccharides; Problem Solving; Production; Property; Proteins; Recombinant Proteins; Recombinants; Research; Safety; Side; Small Business Innovation Research Grant; Structure; System; Testing; Therapeutic; Time; Translating; Vaccines; Validation; cell bank; clinical effect; clinical efficacy; commercial application; commercialization; cost; design; glycosylation; immunogenic; innovation; meetings; public health relevance; sugar; therapeutic protein; tool

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）都是使用哺乳动物细胞系生产的，这些细胞系可以增加其临床功效所需的人型聚糖。但是，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，它们可以包含不定的药物，并且可以产生免疫原性的聚糖。 Baculovirus-insect细胞系统（BICS）提供了一种有吸引力的替代方法，因为它是快速，安全的，并且已经被用于生产两种FDA许可的疫苗。尽管有这些优势，但BIC尚未用于制造非疫苗生物制剂，因为它无法产生其功效所需的人类型n-glycans。我们以前的研究表明，我们可以通过糖化昆虫细胞来解决该问题，以使其N-糖基化途径人性化。但是，这些学术努力并非旨在创建能够以低成本以有效的人型N-糖基化的最佳糖化昆虫细胞系。因此，该实验室对市场项目的总体目标是创建新的糖化昆虫细胞系，这些昆虫细胞系可以提供有效的人类型N-糖基化，而无需昂贵的媒体补充剂以用于商业生物制造和基本生物医学研究。我们将通过在原始，孤立且有据可查的条件下以新的和创新的方式扩展我们以前的努力并以新的和创新的方式扩展我们以前的努力和糖化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，旨在产生与商业生物制剂中通常发现的不同人型N-聚糖的糖蛋白。我们将在原始的，孤立的和有据可查的条件下生产和维持这些细胞系。然后，我们将生产细胞库，对其进行测试是否污染物，并广泛地表征细胞系，以促进未来的验证和商业化工作。我们还将通过将新的糖化昆虫细胞系作为制造平台的可行性，通过将其生产水平与父母细胞系进行比较，使用三种不同的糖蛋白生物制剂与本机N-聚糖结构与与设计相同的天然N-聚糖结构，并通过设计或多个新的昆虫细胞系。最后，我们将通过表达和纯化每种生物学并定量概述其N-聚糖结构来研究我们新昆虫细胞系的人类型N-糖基化效率。 Glycobac，LLC拥有保护糖工程BIC的关键知识产权的独家许可。在创建了本文所述的新细胞系，证明它们的能力并确认其在II阶段中的安全性和功效之后，糖会使这些群体分散 与多个生物技术公司和合同制造组织的界限。 Glycobac还将为全球基本和应用生物医学研究社区提供开放访问这些新细胞系。因此，该项目中产生的工具将对人类医学和基本生物医学研究的不同领域产生重大影响，需要正确和/或可预测的糖基化重组糖蛋白。