Microbioreactor for lower cost and faster optimisation of protein production

Mayur Parekh, Abdulaziz Ali, Zulfiqur Ali, Simon Bateson, Fathi Abugchem, Leon Pybus, Christopher Lennon

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Optimisation of bioprocesses relies on approaches that are either labour intensive or require expensive robotic systems. There is a need for fluidic processing at low volume that can be integrated with existing bioprocess analytics to provide analytical information for the development and optimisation of bioprocesses.
We demonstrate a 1 mL polymer inkjet 3D printed (i3DP) microbioreactor with integrated sensing (pH, oxygen and cell density) for optimisation of recombinant protein production with different feeds. A pressurised fluid driving system was used to control flow rates down to 0.7 μL min−1 with fluid switching from four reservoirs using a manifold controlled by solenoid valves. Oxygen transferred from a headspace via a gas-permeable membrane achieved a kLa of up to 90 h−1 at 1500 rpm. Cultivation of E. coli within the microbioreactor was comparable with a 2 L bench scale bioreactor, with optical densities of respectively 7.1 ± 0.4 and 6.5 ± 0.35. Triplicate batch cultivations within the microbioreactor of Pichia pastoris, with diauxic growth on glycerol (0.20 ± 0.02 h−1) and methanol (0.02 ± 0.04 h−1), showed good control of pH and DO and achieved a maximum dry cell weight of 10 ± 1 g L−1. For continuous cultivations, recombinant protein production was higher in pure methanol (314 ± 23) than methanol-sorbitol (202 ±17) but reduces over time with lower cellular viability for methanol-glucose mixed feed, with less total protein produced and increases in DNA and proteases released. The developed system could be used in different applications including within synthetic biology, cell and gene therapy and organ-on-chips.
Original languageEnglish
Pages (from-to)6148-6161
Number of pages14
Issue number18
Publication statusPublished - 21 Aug 2020

Bibliographical note

Funding Information:
This work was supported by the European Commission through the Horizon 2020 GateOne project (project number 644856) and also by the UKRI, Research England, Connecting Capability Fund through the Thyme project (project code ccf14-7167). MP wishes to acknowledge FUJIFILM Diosynth Biotechnologies for partial support of a studentship and AA wishes to acknowledge support of the Libyan Education Ministry for support of studentship. We wish to thank and remember the many valuable contributions of Dr Simon M. Scott who sadly passed away before preparation of this work.

Publisher Copyright:
© 2020 The Royal Society of Chemistry.

Copyright 2020 Elsevier B.V., All rights reserved.


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