Scalable cell-free manufacturing of diverse therapeutic proteins

Lead Research Organisation: University of Edinburgh
Department Name: Sch of Biological Sciences

Abstract

The aim of this project is to develop cell-free systems which are capable of producing a diverse range of therapeutically-relevant proteins, and investigate how they might be scaled up in an industrial setting. The PhD will be performed in partnership with Fujifilm Diosynth Biotechnologies UK (FDBK).
Cell-free biomanufacturing uses crude cell extracts, or defined systems of purified components, to carry out bioproduction decoupled from cellular growth, which allows efficient channelling of all resources to recombinant protein expression. This could potentially lead to more reproducible production, reduce post-expression modifications which would otherwise lead to product heterogeneity, and reduce levels of cellular-derived contaminants which have to be removed by stringent purification steps. Additionally, cell-free systems may enable the production of cytotoxic proteins which could not be manufactured by whole cell systems. Finally, the lack of a cell wall results in open reactions whose conditions may be adjusted in real time to facilitate protein folding, changing redox environments, and rapid prototyping of complex metabolic pathways.
For these reasons, we propose to investigate the use of cell-free systems to augment or replace existing processes at FDBK, which currently involve the use of microbial and mammalian cells for the production of biotherapeutic recombinant proteins.
The project will be structured around three main objectives:
1. Optimise E. coli lysate system to produce therapeutically-relevant proteins at lab scale, using automation and detailed metabolomic profiling to engineer extracts capable of producing all required proteins with high yield and quality.
2. Develop reconstituted cell-free system with economically competitive performance, using techniques mastered in the first objective.
3. Tech transfer and beyond lab-scale reactions, using FDBK facilities to operate reactions up to 100L-scale. This will demonstrate successful tech transfer to an industrial setting, and result in an informed cost-benefit analysis for cell-free production at industrial scales.
The student will join a highly collaborative lab within the prestigious Centre for Synthetic and Systems Biology (SynthSys), and will be encouraged to make full use of the expertise and world-class facilities available. He/she will gain an in-depth training in relevant experimental techniques for cell-free research, including molecular biology, metabolomics, microscopy, and microfluidics.
Access will be given to the student to the upstream and downstream facilities at FDBK in the North of England during the studentship. This will enable the relevance of the work in an industrial environment to be seen, and also provide the student with an insight into the operations of a technology driven Contract and Development and Manufacturing Organisation within the biotherapeutic sector. The student will also be part of a cohort of students in the FBDK Centre of Excellence in BioProcessing 2.0, and as such will participate in technical meetings at FDBK. The opportunity to immerse themselves in the FDBK culture will give the student a valuable insight into the mechanisms and challenges of industrial biotechnology.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/T00875X/1 01/10/2020 30/09/2028
2435341 Studentship BB/T00875X/1 01/10/2020 30/09/2024 Alex John Perkins