13 ERA IB: Investigating NOvel VAluable bio-Therapeutics and Expression systems

IgG的生产已经彻底改变了生物技术产业，其产品用于治疗癌症和免疫紊乱。经过多年的优化，标准IgG抗体获得了成功，这使得人们对新型重组蛋白（rp）——“第三代”生物制药寄予了很高的期望。时间框架、利益相关者和商业压力与十年前不同，新rp的制造具有挑战性。新的rp如融合蛋白（用于新型疾病靶向）、IgA（最丰富的igg亚型，用于治疗IgA缺乏和癌症）和分泌粘蛋白（用于疾病治疗，疫苗）的商业和临床回报的前景是显而易见的，但这些产品的生产出乎意料地困难，无法预测产生任何（或足够的）产品的失败。可获得的研究表明，在翻译、折叠和翻译后加工（特别是N-和o -糖基化）的水平上存在不可预测的限制。该项目将开发新的动物（CHO）和植物（烟草）细胞表达技术，并建立生物工艺，以生产难以表达的新型rp。目标是(1)确定与难以表达的rp产生相关的分子事件；(2)设计新的表达技术，通过操纵翻译和分泌机制来增强这些分子的合成、折叠和组装；(3)对CHO细胞系进行工程改造，实现IgAs的N-和o -糖基化；(4)设计并建立能够通过定制的糖基化生产工业相关数量的IgAs的植物细胞系；(5)了解表达挑战性rPs的细胞的代谢负荷；(6)建立新的表达技术和rp的上下游流程，并评估扩大规模的潜力。该联盟将为新的表达技术寻求知识产权保护，因为这些技术将对工业界产生巨大的兴趣。

The production of IgG's has revolutionized the biotech industry with products used for the treatment of cancer and immune disorders. The success of standard IgG's built over years of optimization, has led to high expectations for novel recombinant proteins (rPs), "third generation" biopharmaceuticals. Timeframes, stakeholders and commercial pressures are different to those of a decade ago and the manufacture of new rPs is challenging. The promise of commercial and clinical returns from new rPs such as fusion proteins (for novel disease targeting), IgA (the most abundant Ig isoform, for treatment of IgA deficiencies, cancer) and secreted mucins (for disease therapy, vaccines) is obvious but these are unexpectedly difficult to manufacture, with unpredictable failures to generate any (or enough) product. Studies accessible to scrutiny indicate unpredictable limitations occur at the level of translation, folding and post-translational processing (especially N- and O-glycosylation). This project will develop new animal (CHO) and plant (tobacco) cell expression technologies, and establish bioprocesses, for the production of difficult to express and novel rPs. Objectives are to (1) define the molecular events linked to production of difficult to express rPs; (2) engineer new expression technologies for enhanced synthesis, folding and assembly of such molecules via manipulation of translational and secretory machinery; (3) engineer CHO cell lines for the efficent N- and O-glycosylation of IgAs; (4) engineer and establish plant cell lines capable of producing industrially relevant amounts of IgAs with bespoke glycosylation; (5) develop an understanding of the metabolic load on the cell expressing challenging rPs; and (6) establish upstream and downstream processes for the new expression technologies and rPs and evaluate the potential for scale-up. The consortium will seek protection of IP for the new expression technologies, as these will be of immense interest to industry.