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Evaluating and mitigating clinical samples matrix effects on TX-TL cell-free performance

View ORCID ProfilePeter L. Voyvodic, Ismael Conejero, Khouloud Mesmoudi, View ORCID ProfileEric Renard, View ORCID ProfilePhilippe Courtet, View ORCID ProfileDiego I. Cattoni, View ORCID ProfileJerome Bonnet
doi: https://doi.org/10.1101/2022.05.02.489947
Peter L. Voyvodic
1Centre de Biologie Structurale (CBS). INSERM U1054, CNRS UMR5048, University of Montpellier, France
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Ismael Conejero
1Centre de Biologie Structurale (CBS). INSERM U1054, CNRS UMR5048, University of Montpellier, France
2Department of Psychiatry, CHU Nîmes, University of Montpellier, Nîmes, France
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Khouloud Mesmoudi
1Centre de Biologie Structurale (CBS). INSERM U1054, CNRS UMR5048, University of Montpellier, France
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Eric Renard
3UMR CNRS 5203/INSERM U1191, Institute of Functional Genomics, University of Montpellier, Montpellier, 34090, France
4Department of Endocrinology, Diabetes, Nutrition and CIC INSERM 1411, Montpellier University Hospital, Montpellier, 34295, France
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Philippe Courtet
5CHU Montpellier, Department of Emergency Psychiatry and Post Acute Care, Lapeyronie Hospital, CHRU Montpellier, Montpellier, 34090, France
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Diego I. Cattoni
1Centre de Biologie Structurale (CBS). INSERM U1054, CNRS UMR5048, University of Montpellier, France
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  • For correspondence: jerome.bonnet@inserm.fr diego.cattoni@inserm.fr
Jerome Bonnet
1Centre de Biologie Structurale (CBS). INSERM U1054, CNRS UMR5048, University of Montpellier, France
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  • For correspondence: jerome.bonnet@inserm.fr diego.cattoni@inserm.fr
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Abstract

Cell-free biosensors are promising tools for medical diagnostics, yet their performance can be affected by matrix effects arising from the sample itself or from external components. Here we systematically evaluate the performance and robustness of cell-free systems in serum, plasma, urine, and saliva using two reporter systems, sGFP and luciferase. In all cases, clinical samples have a strong inhibitory effect. Of different inhibitors, only the RNase inhibitor mitigated matrix effects. However, we found that the recovery potential of RNase inhibitor was partially muted by interference from glycerol contained in the commercial buffer. We solved this issue by designing a strain producing an RNase inhibitor protein requiring no additional step in extract preparation. Furthermore, our new extract yielded higher reporter levels than previous conditions and tempered interpatient variability associated with matrix effects. This systematic evaluation and improvements of cell-free system robustness unified across many types of clinical samples is a significant step towards developing cell-free diagnostics for a wide range of conditions.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 02, 2022.
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Evaluating and mitigating clinical samples matrix effects on TX-TL cell-free performance
Peter L. Voyvodic, Ismael Conejero, Khouloud Mesmoudi, Eric Renard, Philippe Courtet, Diego I. Cattoni, Jerome Bonnet
bioRxiv 2022.05.02.489947; doi: https://doi.org/10.1101/2022.05.02.489947
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Evaluating and mitigating clinical samples matrix effects on TX-TL cell-free performance
Peter L. Voyvodic, Ismael Conejero, Khouloud Mesmoudi, Eric Renard, Philippe Courtet, Diego I. Cattoni, Jerome Bonnet
bioRxiv 2022.05.02.489947; doi: https://doi.org/10.1101/2022.05.02.489947

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