Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Bernard Katz Building, Gordon Street, London WC1H 0AH, United Kingdom.
Biotechnol J. 2018 Mar;13(3):e1700390. doi: 10.1002/biot.201700390. Epub 2017 Dec 5.
The High Throughput (HT) investigation of chromatographic separations is an important element of downstream bioprocess development due to the importance of chromatography as a technique for achieving stringent regulatory requirements on product purity. Various HT formats for chromatography exist, but the miniature column approach has characteristics resembling large scale packed bed column chromatography the most. The operation of such columns on robotic stations can be automated, but this is not always a straightforward procedure; the robotic manipulations are highly dependent on the settings of each experiment and the standard commands of the supporting software may not provide readily the required flexibility and accessibility for "plug and play" functionality. These can limit the potential of this technique in laboratories engaging on HT activities. In this work, we present an application which aims to overcome this challenge by providing end-users with a flexible operation of the miniature column technique on an automated liquid handler. The application includes a script which is written on Freedom EVOware, and is supplemented by custom compiled executables. Here, the manipulations carried out by the application are described in detail and its functionality is demonstrated through typical experiments based on bind and elute miniature column chromatography. The application is shown to allow for the unsupervised "on-the-fly" programming of the robotic station and to ultimately make the technique accessible to non-automation experts. This application is therefore well suited to simplifying development activities based on the robotic deployment of the miniature column chromatography technique.
高通量（HT）色谱分离研究是下游生物工艺开发的重要组成部分，因为色谱技术对于实现产品纯度严格的监管要求非常重要。存在各种 HT 格式的色谱，但微型柱方法与大规模填充床柱色谱最为相似。这种柱子在机器人站上的操作可以自动化，但这并不总是一个简单的过程；机器人操作高度依赖于每个实验的设置，并且支持软件的标准命令可能无法为“即插即用”功能提供所需的灵活性和可访问性。这些可能会限制参与 HT 活动的实验室对这项技术的潜在应用。在这项工作中，我们提出了一种应用程序，旨在通过为用户提供在自动化液体处理站上灵活操作微型柱技术来克服这一挑战。该应用程序包括一个在 Freedom EVOware 上编写的脚本，并辅以自定义编译的可执行文件。这里详细描述了应用程序执行的操作，并通过基于绑定和洗脱微型柱色谱的典型实验展示了其功能。该应用程序被证明允许对机器人站进行无人监督的“实时”编程，并最终使非自动化专家也能够使用该技术。因此，该应用程序非常适合简化基于微型柱色谱技术的机器人部署的开发活动。
