Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis
Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis
复制标题
通过无细胞蛋白质合成实现护理点肽激素生产
DOI：
10.1021/acssynbio.2c00680
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发表时间：
2023
影响因子：
4.7
通讯作者：
Jewett, Michael C.
中科院分区：
文献类型：
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作者：
DeWinter, Madison A.;Thames, Ariel Helms;Guerrero, Laura;Kightlinger, Weston;Karim, Ashty S.;Jewett, Michael C.
在资源有限的环境中，由于冷链和基础设施的限制，很难将敏感的生物药物安全地提供给患者。护理点药物制造可以规避这些挑战，因为药物可以在当地生产并按需使用。为了实现这一愿景，我们将联合收割机无细胞蛋白质合成（CFPS）和2合1亲和纯化和酶切方案相结合，开发了一个用于即时药物制造的平台。作为一个模型，我们使用这个平台来合成一组肽激素，这是一类重要的药物，可用于治疗各种疾病，包括糖尿病，骨质疏松症和生长障碍。通过这种方法，当需要时，温度稳定的冻干CFPS反应组分可以与编码SUMO化的感兴趣的肽激素的DNA再水化。Strep-Tactin亲和纯化和珠上SUMO蛋白酶切割产生其天然形式的肽激素，其被ELISA抗体识别并且可以结合其各自的受体。随着进一步的发展，以确保适当的生物活性和患者安全，我们设想这个平台可以用于以分散的方式制造有价值的肽激素药物。
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. Toward 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 affinity purification and on-bead SUMO protease cleavage yield 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 that this platform could be used to manufacture valuable peptide hormone drugs in a decentralized way.