Flexible and Accessible Automated Operation of Miniature Chromatography Columns on a Liquid Handling Station

Abstract

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.

Microbioreactor Systems for Accelerated Bioprocess Development

“…In the past, the integration of robotic systems for downstream process development led to significant increases in the throughput of several standard operation procedures. For example, cell lysis has been optimized for improved release of intracellular products, the screening of chromatographic material and binding/elution conditions was facilitated, and the identification of optimal compositions of aqueous two‐phase systems or precipitation conditions has become experimentally feasible. So far only a few but highly interesting studies have been reported that combine high‐throughput upstream and downstream bioprocess development .…”

Holistic process development to mitigate proteolysis of a subunit rotavirus vaccine candidate produced in Pichia pastoris by means of an acid pH pulse during fed‐batch fermentation

“…(a) Fermentations carried out at the indicated pH, (b) fermentations carried out with a pH pulse or pH shift, (c) scale for P2-VP8-P [4] concentration, the area of the circle showing full-length and truncated P2-VP8-P [4] species (a and b) is proportional to its concentration in the supernatant 2 or 20 μL per well (GE Healthcare Life Sciences) 27 or 0.6 mL RoboColumns (custom packed by Repligen Europe B.V, Breda, The Netherlands). 28 A total of 18 resins were screened for binding capacity and product quality (Table 1)…”

High-Throughput Process Development for the Chromatographic Purification of Viral Antigens

“…While batch HT methods allow the evaluation of multiple conditions in parallel, including binding capacity measurements from small product masses due to the low volumes of stationary phases employed. Moreover, the miniature packed bed column technique (i.e., RoboColumns) can be easily scaled-up or -down [7]. It can also return an efficient way of evaluating different stationary phases due to the advantage of packing any beaded based resin at a cost.…”

“…The followed workflow employs a Tecan Freedom EVO® 200 automated liquid handling station (Tecan Group Ltd., Männedorf, Switzerland) and MATLAB® (The MathWorks, MA, USA) codes, compiled into executables, which convert user-defined inputs into robotic commands. These are then executed within generic Tecan Freedom EVOware® (Tecan Group Ltd.) based scripts that implement all necessary actions to complete a study with no end-user intervention [7]. Here, both 0.2 and 0.6 mL columns, packed with various types of resin, can be used in flowthrough or bind and elute mode, with the latter employing isocratic, multi-step and multi-slope gradient elution.…”