Point-of-care peptide hormone production enabled by cell-free protein synthesis
DeWinter, M. A.; Thames, A. H.; Guerrero, L.; Kightlinger, W. K.; Karim, A. S.; Jewett, M. C.
Show abstract
In resource-limited settings, it can be difficult to safely deliver sensitive biologic medicines to patients due to cold chain and infrastructure constraints. Point-of-care drug manufacturing could circumvent these challenges since medicines could be produced locally and used on-demand. Towards this vision, we combine cell-free protein synthesis (CFPS) and a 2-in-1 affinity purification and enzymatic cleavage scheme to develop a platform for point-of-care drug manufacturing. As a model, we use this platform to synthesize a panel of peptide hormones, an important class of medications that can be used to treat a wide variety of diseases including diabetes, osteoporosis, and growth disorders. With this approach, temperature-stable lyophilized CFPS reaction components can be rehydrated with DNA encoding a SUMOylated peptide hormone of interest when needed. Strep-Tactin(R) affinity purification and on-bead SUMO protease cleavage yields peptide hormones in their native form that are recognized by ELISA antibodies and that can bind their respective receptors. With further development to ensure proper biologic activity and patient safety, we envision this platform could be used to manufacture valuable peptide hormone drugs at the point-of-care in resource-limited settings.
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