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Development of a high-throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design

Bláha, BAF; Morris, SA; Ogonah, OW; Crescente, V; Rosenberg, W; Maucourant, S; Mukhopadhyay, TK; (2018) Development of a high-throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design. Biotechnology Progress , 34 (1) pp. 130-140. 10.1002/btpr.2555. Green open access

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Abstract

The time and cost benefits of miniaturized fermentation platforms can only be gained by employing complementary techniques facilitating high-throughput at small sample volumes. Microbial cell disruption is a major bottleneck in experimental throughput and is often restricted to large processing volumes. Moreover, for rigid yeast species such as Pichia pastoris, no effective high-throughput disruption methods exist. This study describes the development of an automated, miniaturized, high-throughput, non-contact, scalable platform based on Adaptive Focused Acoustics (AFA) to disrupt P. pastoris and recover intracellular heterologous protein. Augmented modes of AFA were established by investigating vessel designs and a novel enzymatic pre-treatment step. Three different modes of AFA were studied and compared to the performance high pressure homogenization. For each of these modes of cell disruption, response models were developed to account for five different performance criteria. Using multiple responses not only demonstrated that different operating parameters are required for different response optima, with highest product purity requiring suboptimal values for other criteria, but also allowed for AFA-based methods to mimic large-scale homogenization processes. These results demonstrate that AFA-mediated cell disruption can be used for a wide range of applications including buffer development, strain selection, fermentation process development and whole bioprocess integration. This article is protected by copyright. All rights reserved.

Type: Article
Title: Development of a high-throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.2555
Publisher version: http://dx.doi.org/10.1002/btpr.2555
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Adaptive Focused Acoustics, Pichia pastoris, cell disruption, high-throughput, homogenization, scale-down
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Inst for Liver and Digestive Hlth
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: http://discovery.ucl.ac.uk/id/eprint/1573123
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