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

Abstract

Chromatography is a widely used method in the biotechnology industry, it functions to separate the desired product from process and product related impurities. There is a multitude of resins available based on different modalities (such as charge, hydrophobicity and affinity) to provide a spectrum of approaches to meet the separation challenges of the diverse products. The challenge of developing viral antigen purification processes is addressed in this method. A unique feature of this product class is that in order to protect against more than one strain of an antigen, vaccines are often multivalent. This entails multiple production processes for each antigen, all of which will require separate development and validation.Ideally, a universal purification method is sought, but differences in the protein subunits (frequently used as the antigens) makes this challenging and often bespoke purification steps are required. This means process development for the chromatographic stages of these products can be particularly challenging and labour intensive. With the numerous choices available, making critical process decisions that are usually unique to each product, process and strain, can be costly and time-consuming. To address this, scale down purification at < 1.0 mL column volume and automation approaches are increasingly applied to increase throughput. In this work, a method is described wherein a Tecan Freedom EVO ® automated liquid handler is deployed for the evaluation of different resin chemistries and buffer conditions to find a suitable purification strategy. This method allows for the rapid evaluation of the separation viral antigens where limited information on chromatography behaviour is known at the early stages of process development. Here, we demonstrate the methodology firstly by explaining the automated purification script and secondly by applying the script for an efficient purification development for different serotypes of rotavirus antigens.

Automated liquid-handling operations for robust, resilient, and efficient bio-based laboratory practices

“…Moreover, the inclusion and utilisation of a liquid handling device was decided on the basis that it helped to solve a specific problem or overcome certain challenges. Most of the applications shown in Table 2 are related to the ability to process a large number of samples in a short period of time [21][22][23][24][25][26][27][28][29][30][31]. This is one of the most evident technical advantages of a liquid handling device to be used for the execution of a simple task, i.e., dispensing of some liquid into a specific location, which can become time-consuming when performed at scale.…”

“…The use of multi-channel pipetting with precise flow control has allowed scale-down chromatographic experiments to be performed. Miniaturized columns have not only allowed to accelerate the discovery of adequate purification conditions for pharmaceutical products [21,33], but also to identify key bottlenecks and obstacles before performing large-scale experimentation [34]. Cartridge-based purification has also been applied for solid-phase extraction [33], enrichment of phosphorylated proteins [35] and quantification of metabolites in clinical samples [36].…”

High throughput chromatography and analytics can inform viral clearance capabilities during downstream process development for biologics

“…When HTS is performed using these miniature columns in conjunction with a liquid handling device, the results prove scalable and predictive of lab‐scale processes, primarily due to representative packed bed characteristics and well‐controlled flow rates. [ 5–8 ] Thus, HTS has become an invaluable tool for identifying and selecting optimal process conditions early in the development timeline.…”