Pilot Production of SARS-CoV-2 Related Proteins in Plants: A Proof of Concept for Rapid Repurposing of Indoor Farms Into Biomanufacturing Facilities
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Abstract
The current CoVid-19 crisis is revealing the strengths and the weaknesses of the world’s capacity to respond to a global health crisis. A critical weakness has resulted from the excessive centralization of the current biomanufacturing capacities, a matter of great concern, if not a source of nationalistic tensions. On the positive side, scientific data and information have been shared at an unprecedented speed fuelled by the preprint phenomena, and this has considerably strengthened our ability to develop new technology-based solutions. In this work, we explore how, in a context of rapid exchange of scientific information, plant biofactories can serve as a rapid and easily adaptable solution for local manufacturing of bioreagents, more specifically recombinant antibodies. For this purpose, we tested our ability to produce, in the framework of an academic lab and in a matter of weeks, milligram amounts of six different recombinant monoclonal antibodies against SARS-CoV-2 in Nicotiana benthamiana . For the design of the antibodies, we took advantage, among other data sources, of the DNA sequence information made rapidly available by other groups in preprint publications. mAbs were engineered as single-chain fragments fused to a human gamma Fc and transiently expressed using a viral vector. In parallel, we also produced the recombinant SARS-CoV-2 N protein and the receptor binding domain (RBD) of the Spike protein in planta and used them to test the binding specificity of the recombinant mAbs. Finally, for two of the antibodies, we assayed a simple scale-up production protocol based on the extraction of apoplastic fluid. Our results indicate that gram amounts of anti-SARS-CoV-2 antibodies could be easily produced in little more than 6 weeks in repurposed greenhouses with little infrastructure requirements using N. benthamiana as production platform. Similar procedures could be easily deployed to produce diagnostic reagents and, eventually, could be adapted for rapid therapeutic responses.
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SciScore for 10.1101/2020.10.13.331306: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Cloning of anti-SARS-CoV-2 antibodies and SARS-CoV-2 antigens: All sequences were cloned and assembled using the GoldenBraid (GB) assembly system (Sarrion-Perdigones et al., 2013; https://gbcloning.upv.es). anti-SARS-CoV-2suggested: NoneAll antibodies were cloned as single chain antibodies fused to the human IgG1 Fc domain. human IgG1 Fc domain.suggested: NoneFor anti-SARS-CoV-2 antibody detection, the blots were incubated with 1:20000 HRP-conjugated rabbit anti-human IgG (Sigma-Aldrich, … SciScore for 10.1101/2020.10.13.331306: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Cloning of anti-SARS-CoV-2 antibodies and SARS-CoV-2 antigens: All sequences were cloned and assembled using the GoldenBraid (GB) assembly system (Sarrion-Perdigones et al., 2013; https://gbcloning.upv.es). anti-SARS-CoV-2suggested: NoneAll antibodies were cloned as single chain antibodies fused to the human IgG1 Fc domain. human IgG1 Fc domain.suggested: NoneFor anti-SARS-CoV-2 antibody detection, the blots were incubated with 1:20000 HRP-conjugated rabbit anti-human IgG (Sigma-Aldrich, #A8792). anti-human IgGsuggested: (Sigma-Aldrich Cat# A8792, RRID:AB_258414)For SARS-CoV-2 antigen detection the blots were incubated with 1:2000 Anti-His mouse monoclonal primary antibody (Qiagen, #34660) and then incubated with 1:10000 peroxidase labelled anti-mouse IgG secondary antibody (GE Healthcare). Anti-Hissuggested: Noneanti-mouse IgGsuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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