Development of an Improved Bio-Manufacturing Platform for the Production of Glyco

开发用于生产糖的改进生物制造平台

• 批准号: 8780700
• 负责人: Beth Rasala
• 金额: $ 12.31万
• 依托单位: TRITON ALGAE INNOVATIONS, INC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8780700
• 关键词: Address; Age; Algae; Animals; Biological; Biological Assay; Biological Products; Cell Culture Techniques; Cell Line; Cells; Chlamydomonas; Chlamydomonas reinhardtii; Collaborations; Complex; Computer Simulation; Data; Development; Drug Industry; EST Library; Engineering; Enzymes; Erythropoietin; Eukaryota; Genome; Glycobiology; Glycoproteins; Glycoside Hydrolases; Green Algae; Half-Life; Health; Heterogeneity; Human; Insecta; Investigation; Legal patent; Light; Link; MALDI-TOF Mass Spectrometry; Mammals; Marketing; Mass Spectrum Analysis; Nutrient; Pattern; Performance; Pharmacologic Substance; Phase; Plants; Polysaccharides; Price; Production; Productivity; Proteins; Recombinant Proteins; Recombinants; Research; Safety; Series; Serum; Site; Solubility; Structure; System; Temperature; Therapeutic; Therapeutic Uses; Triton; Tritons; Validation; Vascular Plant; Yeasts; cost; gel mobility shift assay; genetic manipulation; glycosylation; glycosyltransferase; immunogenicity; improved; innovation; insight; interest; pressure; protein folding; public health relevance; scale up; therapeutic protein; trafficking

DESCRIPTION (provided by applicant): The protein therapeutic market is valued at $99 billion and is the fastest growing segment of the $600 billion pharmaceutical industry. Nearly half of all approved biopharmaceuticals are post-translationally modified by glycosylation [1]. Glycosylation influences protein solubility, serum half-life, immunogenicity and/or biological activity. Animal cell lines are primarily used for the production of therapeutic glycoproteins, due to their ability to generate products with therapeutically acceptable glycoprofiles. However, animal cell lines have limitations including high production costs, reduced safety, and heterogeneity of glycoforms [2,3]. Glycoform heterogeneity can be found both in the site occupancy of the glycan and the structure of the attached glycan; and is influenced by cell culture parameters such as age, pH, nutrient levels, and temperature [4]. Thus, a major challenge for the production of therapeutic glycoproteins from animal cell lines is glycoprofile consistency, and/or the purification of a desirable glycoform from a heterogeneous mixture [2]. Furthermore, an estimated $26 billion worth of biologics will lose patent protection by 2016, creating significant market opportunities for price-competitive biosimilars [5]. In light of these challenges and predicted market pressures, significant research is ongoing to develop alternative expression platforms for therapeutic glycoproteins with acceptable glycoprofiles. Triton Algae Innovations, Inc. (Triton) is developing a recombinant protein production platform utilizing a photosynthetic singled-celled green alga, Chlamydomonas reinhardtii. There are several advantages to using green microalgae as an expression platform for protein therapeutics: cost, safety, scalability, productivity, ease of genetic manipulation, and ability to produce complex eukaryotic proteins. We have proven the value of this platform with the production of an aglycosylated mammalian protein [6] that will be commercially available next year. Despite these attributes and validation, several questions need to be addressed as the system develops into a bio-manufacturing platform, especially the glycosylation profile of recombinantly expressed human therapeutic proteins. Triton will use a well-characterized and therapeutically valuable protein, human erythropoietin (EPO), to better understand glycan structures attached to recombinant glycoproteins produced by C. reinhardtii.

描述（由申请人提供）：蛋白质治疗市场价值990亿美元，是6000亿美元制药行业中增长最快的部分。所有批准的生物制药中有近一半是通过糖基化进行后修饰的[1]。糖基化影响蛋白质溶解度、血清半衰期、免疫原性和/或生物活性。动物细胞系主要用于生产治疗性糖蛋白， 它们产生具有治疗上可接受的糖谱的产物的能力。然而，动物细胞系具有局限性，包括高生产成本、降低的安全性和糖型的异质性[2，3]。糖型异质性存在于聚糖的位点占有率和连接聚糖的结构中;并受细胞培养参数（如年龄、pH值、营养水平和温度）的影响[4]。因此，从动物细胞系生产治疗性糖蛋白的主要挑战是糖谱的一致性，和/或从异质混合物中纯化所需的糖型[2]。此外，到2016年，估计有价值260亿美元的生物制剂将失去专利保护，这为具有价格竞争力的生物仿制药创造了巨大的市场机会。鉴于这些挑战和预测的市场压力，正在进行重要的研究以开发具有可接受的糖谱的治疗性糖蛋白的替代表达平台。Triton Algae Innovations，Inc.（Triton）正在开发利用光合作用单细胞绿色藻类莱茵衣藻（Chlamyeliumreinhardtii）的重组蛋白生产平台。使用绿色微藻作为蛋白质治疗剂的表达平台有几个优点：成本、安全性、可扩展性、生产率、遗传操作的容易性和表达能力。 产生复杂的真核蛋白质。我们已经证明了这个平台的价值，生产了一种无糖基化的哺乳动物蛋白[6]，将于明年上市。尽管有这些属性和验证，但随着该系统发展成为生物制造平台，需要解决几个问题，特别是重组表达的人类治疗蛋白质的糖基化谱。Triton将使用一种具有良好特征和治疗价值的蛋白质，人促红细胞生成素（EPO），以更好地了解连接到由C.莱因哈德氏菌