A novel screen for product molecule compatibility with the production cell that accounts for intracellular toxicity

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CBMNet Funded Proof-of-Concept project: A novel screen for product molecule compatibility with the production cell that accounts for intracellular toxicity – Dr Jagroop Pandhal, Croda and The Centre for Process Innovation

You can download the University of Sheffield Proof-of-Concept case study

“One of the key challenges for competitive  IB  is to understand the metabolic impact of product and intermediates accumulation arising from  novel inserted synthetic pathways.  This project starts to build out  the necessary toolbox  required for the early characterisation, selection and engineering of the optimum host/constructs necessary for viable production processes.   We therefore applaud The University of Sheffield’s focus in this area and will continue to be supportive of these initiatives. ” – Rob Mitra, The Centre for Process Innovation

The Challenge

Industrial Biotechnology increasingly uses genetically-enhanced production cells to biosynthesise non-native chemical products. Being non-native and high concentration, such products often are toxic to the production cell.

This being so, many putative products applied externally to test for compatibility with the production cell will not cross the membrane, so intracellular cytotoxicity – the key IB question – is not assayed.

The Research

Dr Jagroop Pandhal is a Lecturer in Biological Engineering at the University of Sheffield. The research in his laboratory focusses mainly on metabolic engineering and biomanufacturing, for example, for therapeutic protein production.

Dr Pandhal applied for CBMNet Proof-of- Concept funding with Croda and The Centre for Process Innovation. The project aimed to develop superior cell toxicity screen exploiting a membrane disruption methodology in cells to achieve product uptake.

The Result

The new screen worked. Two anonymous industrial bio-products were tested with both the traditional screen and the new test.

The new screen demonstrated a specific level of toxicity, missed by the present screen, and therefore these findings could link to productivity levels for the compounds.

With this knowledge, the aim would be to improve productivity with process improvements or cell engineering. Moreover, this methodology could be developed for application to different industrially relevant host cell organisms.

The Future

The industrial collaborators are interested in the test for clients who want to test toxicity of products and also the response of cells to inhibitory compounds in complex feedstocks and are looking to progress this very soon.

The data from this project will be used to apply for funding through the Engineering and Physical Sciences Research Council’s Impact, Innovation and Knowledge Exchange Proof of Concept funding scheme to undertake these tests on more molecules. 

‘Toxicity of products is a significant concern when developing products using IB and, especially, Synthetic Biology where the product is non-native and may be new to nature. The approach devised at Sheffield through this project shows significant potential. Availability of a robust test method, which accounts for trafficking across the membrane, represents a valuable tool for industry.’’ – Dr Doug Cossar, Croda

‘’Being able to develop an improved screen within the laboratory was a key aim and interesting to undertake. However, using it to test real-life industrial biotechnology products and discovering potential toxicity, which manufacturers don’t see, was a revelation.” – Dr Jagroop Pandhal