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

描述(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。所有批准的生物药物中，近一半是通过糖基化进行翻译后修饰[1]。糖基化影响蛋白质的溶解性、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。 它们产生治疗上可接受的糖链的产品的能力。然而，动物细胞系具有局限性，包括生产成本高、安全性降低以及糖型的异质性[2，3]。糖型异质性可以在糖链的位置和连接的糖链的结构中发现，并且受到细胞培养参数的影响，如年龄、pH、营养水平和温度[4]。因此，从动物细胞系生产治疗性糖蛋白的一个主要挑战是糖型的一致性，和/或从不同的混合物中提纯所需的糖型[2]。此外，到2016年，估计价值260亿美元的生物制剂将失去专利保护，为具有价格竞争力的生物仿制药创造巨大的市场机会[5]。鉴于这些挑战和预测的市场压力，正在进行重大研究，以开发具有可接受糖谱的治疗性糖蛋白的替代表达平台。Triton藻类创新公司(Triton)正在开发一种利用光合作用单细胞绿藻--莱茵衣藻--生产重组蛋白质的平台。使用绿色微藻作为蛋白质疗法的表达平台有几个优点：成本、安全性、可伸缩性、生产率、易于遗传操作以及能够 产生复杂的真核蛋白质。我们已经通过生产一种糖基化的哺乳动物蛋白质[6]证明了这个平台的价值，该蛋白质将于明年投入商业使用。尽管有这些属性和有效性，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性蛋白的糖基化图谱。Triton将使用一种具有良好特性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莱茵梭菌产生的重组糖蛋白上的糖链结构。

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.