Towards the automated optimisation of protein labelling methods

该项目的目标是开发自主的、自我优化的方法来改造生物制药。这将通过开发用于反应采样的液体处理系统以及用于反应自动分析的软件平台来实现。目的：1.开发基于MS的蛋白质标记数据的自动去卷积、积分和动力学分析的计算平台。2.开发一种液体处理方法，用于蛋白质标记方法的自动反应设置和采样。3.开发一种计算方法，通过将目标1和目标2联系起来，识别和优化用于自动反应优化的反应条件。方法：最初计划的反应优化方法是使用固相萃取方法自动采样和制备固定时间的反应样品。这些样品(96孔板格式)将在自动去卷积、积分和定量之前通过MS直接分析。反应样品数据将自动集成到一个软件包中，用于动力学分析和随后的反应优化，从而在使用最少试剂的情况下获得最高质量的产品。将使用一系列的蛋白质修饰反应，最初的例子是该大学最近开发的基于多肽连接酶的方法的使用。在该项目中，预计该系统将被应用于各种新的变异蛋白质修饰方法，以进一步例证该方法。潜在影响：该项目将开发一种通用平台技术，用于优化蛋白质-小分子结合物的生产。该方法被设计为对蛋白质和小分子靶标都是不可知的，因此将被转移到任何新的生物药物制备方法。使用这种方法的快速优化将降低优化的成本，并在严格定义生产所需的反应参数以及定义可用作知识产权产品规格一部分的属性方面提供经济优势。

The objective of the project is to develop autonomous, self-optimising approaches towards modified biopharmaceuticals. This will be achieved by development of a liquid-handling system for reaction sampling, together with a software platform for automated analysis of reaction.Aims: 1. Develop a computational platform for automated deconvolution, integration and kinetic analysis of MS-based protein-labelling data. 2. Develop a liquid-handling approach for automated reaction set-up and sampling of protein-labelling approaches. 3. Develop a computational approach to identify and optimise reaction conditions for automated reaction optimisation by linking aim 1 and 2. Methodology:The initial planned approach to reaction optimisation is to use solid-phase extraction approaches to automatically sample and prepare fixed-time reaction samples. These samples (in a 96-well plate format) will then be directly analysed via MS before automated deconvolution, integration and quantification. Reaction sample data will be automatically integrated into a software package for kinetic analysis and subsequent optimisation of reactions to obtain the highest quality product while using minimal reagent quantities. A range of protein modification reactions will be used, initially exemplified by the use of recently-developed peptide-ligase-based methodologies at the University. During the project, it is anticipated that the system will be applied to a variety of novel variant protein modification methods to exemplify the approach further. Potential Impact:The project will develop a generic platform technology for the optimisation of protein-small molecule conjugate production. The approach is designed to be agnostic to both the protein and small molecule target and will therefore be transferable to any new biopharmaceutical preparation method. Rapid optimisation using this approach will reduce both the costs of optimisation and also provide economic advantages in terms of tightly defining reaction parameters required for production as well as defining attributes which can be used as part of the product specification for IP purposes.