Kovar Karin, Looser Verena, Hyka Petr, Merseburger Tobias, Meier Christian
Institute of Biotechnology, School of Life Sciences and Facility Management (Department N), Zurich University of Applied Sciences, Grüental, Wädensswil.
Chimia (Aarau). 2010;64(11):813-8. doi: 10.2533/chimia.2010.813.
Health and safety concerns, enhanced quality criteria, and environmental sustainability, have prompted investigations into production using recombinant yeasts as a feasible alternative for isolation of proteins from natural animal or plant sources, as well as for processes utilising either mammalian cell cultures or bacterial systems. An overview of recent research papers and review articles provides readers with a comprehensive insight into the field of next-generation yeast expression systems. Major breakthroughs in recombinant yeast technology linked to Pichia pastoris are (i) the public availability of tools to generate proteins with tailored and highly homogenous N-glycan structures, similar to the forms assembled in humans, (ii) the recent accomplishment of the annotation of its genome sequence, and finally, (iii) the presence of the first few (non-glycosylated) therapeutic proteins in Pichia on the market. The P. pastoris expression platform is now well developed, as proven by multiple products used in human and veterinary medicine and in industry (e.g., enzymes for chemical synthesis and for the modification/synthesis of pharmaceuticals, drug target proteins used for structural analysis or for high throughput screening, proteins for diagnostics, proteinous biomaterials, vaccines, and therapeutic proteins). Nevertheless, the complexity of protein analysis (monitoring) continues to restrict process development for recombinant products. Drawing on combined expertise in molecular biology and process technology, the Institute of Biotechnology (IBT) at the Zurich University of Applied Science (ZHAW) and its international partners have developed solutions which (i) fully eliminate (or partially reduce) the use of methanol, which is undesirable in high-cell-density and high-productivity processes, (ii) match both strain construction and process design with the target protein characteristics to the benefit of the cells' physiological shape, and (iii) allow multi-gene expressions to be balanced to achieve custom tailored and reproducible protein quality at the level of (engineered) posttranslational modifications. In addition to enabling superior product quality specifications to be achieved with reduced development time, these innovations have helped the industries involved to minimise financial risks and the risk of failure, as well as create an opportunity for (new) drugs with improved functionality at low cost.
健康与安全问题、更高的质量标准以及环境可持续性，促使人们对使用重组酵母进行生产展开研究，这是从天然动物或植物来源分离蛋白质以及利用哺乳动物细胞培养或细菌系统的可行替代方案。近期研究论文和综述文章的概述为读者提供了对下一代酵母表达系统领域的全面洞察。与巴斯德毕赤酵母相关的重组酵母技术的重大突破包括：（i）公开可用的工具可生成具有定制且高度同质的N -聚糖结构的蛋白质，类似于在人类中组装的形式；（ii）最近完成了其基因组序列的注释；最后，（iii）市场上出现了巴斯德毕赤酵母中首批（非糖基化）治疗性蛋白质。巴斯德毕赤酵母表达平台现已得到充分发展，用于人类和兽医学以及工业的多种产品（例如用于化学合成以及药物修饰/合成的酶、用于结构分析或高通量筛选的药物靶蛋白、诊断用蛋白质、蛋白质生物材料、疫苗和治疗性蛋白质）证明了这一点。然而，蛋白质分析（监测）的复杂性仍然限制着重组产品的工艺开发。凭借分子生物学和工艺技术方面的综合专业知识，苏黎世应用科学大学（ZHAW）的生物技术研究所（IBT）及其国际合作伙伴开发了一些解决方案，这些方案（i）完全消除（或部分减少）甲醇的使用，甲醇在高细胞密度和高生产率过程中是不理想的；（ii）使菌株构建和工艺设计与目标蛋白质特性相匹配，以利于细胞的生理形态；（iii）允许平衡多基因表达，以在（工程化的）翻译后修饰水平实现定制且可重复的蛋白质质量。除了能够在缩短开发时间的情况下实现卓越的产品质量规格外，这些创新还帮助相关行业将财务风险和失败风险降至最低，并为以低成本开发具有改进功能的（新型）药物创造了机会。
