Development of workflows for O-glycan profiling and structural characterisation of biopharmaceuticals

2016年，收入排名前十的药物中有一半以上是生物制剂，其中大部分是糖蛋白。因此，糖蛋白疗法对制药业的价值是显而易见的，但这也带来了一个问题。这些药物的生物生产可产生批次间差异性，特别是由于聚糖的非模板生物合成。聚糖对药物功能具有重要的调节作用，并且它们是确定血清半衰期的基础。为了保持药物的安全性和有效性，需要用于聚糖分析的稳健的质量控制方法。此外，需要细胞系工程化以限制聚糖异质性并将生物合成转向有益结构。聚糖分析现在是N-聚糖分析的常规方法，但用于分析第二类主要蛋白质连接聚糖O-聚糖的方法远远落后，并且通常被认为不适合工业采用。需要对O-聚糖进行常规表征和/或定量，以进一步探索与O-聚糖结构变化的功能联系。在Mesquas-Oates（质谱法）和Ungar（细胞生物学）组中，先前共享的博士生已经开发了用于培养细胞的稳健的组合N-和0-聚糖谱分析方法（Abdul Rahman等人，J. Proteome Res.，2014，13：1167-1176; Skeene等人，Anal Chem 2017，89：5840-5849）。在这个BBSRC/IBioIC博士项目中，学生将加入这些小组，在那里他们将建立在以前的工作基础上，为基于糖蛋白的生物制剂生成这样一种集成方法，该方法可以由CRO（如工业合作伙伴Covance）常规使用，以支持未来的药物开发项目。同时，将产生具有改变的O-糖基化的细胞系来挑战新方法，同时还产生具有有益糖基化特性的细胞。科文斯的研发实验室靠近约克，便于频繁的面对面访问。学生每年将在Covance工作约一个月，Covance将提供专业知识，帮助制定拟议的分析工作流程，以满足行业和监管要求，并从项目一开始就提供适当的生物制药材料。通过该计划，学生将获得分析碳水化合物化学，最先进的质谱，糖生物学，细胞生物学和工业生物技术方法的经验，提高他们的就业能力。

In 2016, more than half of the top ten drugs by revenue were biologics, the majority of which are glycoproteins. The value of glycoprotein therapeutics for the pharmaceutical industry is thus clear, but this comes with a catch. The biological production of these drugs can generate batch-to-batch variability, in particular due to the non-templated biosynthesis of glycans. The glycans have important modulating effects on drug function, and they are fundamental for determining serum half-life. In order to maintain drug safety and efficacy, robust quality control methods for glycan analysis are required. In addition, cell line engineering to limit glycan heterogeneity and steer biosynthesis towards beneficial structures is needed. Glycan analysis is now routine for N-glycan profiling but methods for analysis of the second main class of protein-linked glycans, O-glycans, lag well behind, and are generally considered inadequate for industrial adoption. Routine characterisation and/or quantification of O-glycans is required to further explore the functional link with changes in O-glycan structures. In the Thomas-Oates (mass spectrometry) and Ungar (cell biology) groups, previous shared PhD students have developed a robust combined N- and O-glycan profiling method for cultured cells (Abdul Rahman et al, J. Proteome Res., 2014, 13: 1167-1176; Skeene et al., Anal Chem 2017, 89: 5840-5849). In this BBSRC/IBioIC PhD project the student will join these groups, where they will build on the previous work to generate such an integrated method for glycoprotein-based biologics, which can be routinely used by CROs like industrial partner Covance, in support of future drug development projects. At the same time, cell lines with altered O-glycosylation will be generated to challenge the new method, while also producing cells with beneficial glycosylation properties. The location of Covance's R&D laboratories close to York facilitates frequent face-to-face visits. The student will spend approximately one month every year at Covance, and Covance will provide expertise to help shape the proposed analytical workflows to meet industry and regulatory requirements and offer appropriate biopharmaceutical materials from the outset of the project. Through the programme, the student will gain experience in analytical carbohydrate chemistry, state of the art mass spectrometry, glycobiology, cell biology and industrial biotechnology approaches, enhancing their employability.