An integrated robotic system for high‐throughput process development of cell and virus culture conditions: Application to biosafety level 2 live virus vaccines

Daniels, Christopher L.; Rodriguez, Jason; Lim, Esther; Wenger, Marc

doi:10.1002/elsc.201400245

Cited by 7 publications

(3 citation statements)

References 30 publications

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“…Integrated robotic systems offer viable solutions towards enabling full automation across multiple complex processes, such as cell culture, liquid handling and, ultimately, fixation, immunostaining, and plate imaging. These systems have been documented to expedite diverse projects within the pharmaceutical industry ( 32 , 33 ). Nevertheless, to the best of our understanding, there is currently no published study that has applied the full-automation approach to potency measurement of LVVs.…”

Section: Introductionmentioning

confidence: 99%

Fully automated high-throughput immuno-µPlaque assay for live-attenuated tetravalent dengue vaccine development

Wang,

Troutman,

Hofmann

et al. 2024

Front. Immunol.

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Dengue fever has remained a continuing global medical threat that impacts half of the world’s population. Developing a highly effective dengue vaccine, with live-attenuated tetravalent vaccines as leading candidates, remains essential in preventing this disease. For the development of live virus vaccines (LVVs), potency measurements play a vital role in quantifying the active components of vaccine drug substance as well as drug product during various stages of research, development, and post-licensure evaluations. Traditional plaque-based assays are one of the most common potency test methods, but they generally take up to weeks to complete. Less labor and time-intensive potency assays are thus called for to aid in the acceleration of vaccine development, especially for multivalent LVVs. Here, we introduce a fully automated, 96-well format µPlaque assay that has been optimized as a high-throughput tool to evaluate process and formulation development of a live-attenuated tetravalent dengue vaccine. To the best of our knowledge, this is the first report of a miniaturized viral plaque method for dengue with full automation via an integrated robotic system. Compared to the traditional manual plaque assay, this newly developed method substantially reduces testing time by approximately half and allows for the evaluation of over ten times more samples per run. The fully automated workflow, from cell culture to plaque counting, significantly minimizes analyst hands-on time and improves assay repeatability. The study presents a pioneering solution for the rapid measurement of LVV viral titers, offering promising prospects for advancing vaccine development through high-throughput analytics.

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Section: Introductionmentioning

confidence: 99%

Fully automated high-throughput immuno-µPlaque assay for live-attenuated tetravalent dengue vaccine development

Wang,

Troutman,

Hofmann

et al. 2024

Front. Immunol.

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“…High-Throughput (HT) development activities are currently performed in both upstream and downstream unit operations and benefit by the advancements of automated liquid handlers [1, 2,15]. Such screening activities, even from the early stages of process development, can lead to optimal or near optimal conditions in a systematic and efficient fashion [8,9,13,14].…”

Section: Introductionmentioning

confidence: 99%

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

Jacob

Konstantinidis

Bracewell

2020

Vaccine Delivery Technology

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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.

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“…This is a property that can assist the development of a chromatography‐based unit operation by integrating, for example, experiments at micro‐ and bench‐scales. While automation offers a route for achieving increased productivity (e.g.,), the development of a robust and generic automated process is an often non‐trivial task (e.g.,). Within the context of HT bioprocess development the accrued benefits are, however, significant.…”

Section: Introductionmentioning

confidence: 99%

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

Konstantinidis

Goh

Bufájer

et al. 2017

Biotechnology Journal

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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.

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An integrated robotic system for high‐throughput process development of cell and virus culture conditions: Application to biosafety level 2 live virus vaccines

Cited by 7 publications

References 30 publications

Fully automated high-throughput immuno-µPlaque assay for live-attenuated tetravalent dengue vaccine development

Fully automated high-throughput immuno-µPlaque assay for live-attenuated tetravalent dengue vaccine development

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

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

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