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点击电化学用于快速标记病毒、细菌和细胞表面。

Click-electrochemistry for the rapid labeling of virus, bacteria and cell surfaces.

机构信息

Nantes Université, CNRS, CEISAM UMR 6230, F-44000, Nantes, France.

Nantes Université, TaRGeT, Translational Research for Gene Therapies, CHU Nantes, INSERM, UMR 1089, F-44000, Nantes, France.

出版信息

Nat Commun. 2023 Aug 23;14(1):5122. doi: 10.1038/s41467-023-40534-0.

Abstract

Methods for direct covalent ligation of microorganism surfaces remain poorly reported, and mostly based on metabolic engineering for bacteria and cells functionalization. While effective, a faster method avoiding the bio-incorporation step would be highly complementary. Here, we used N-methylluminol (NML), a fully tyrosine-selective protein anchoring group after one-electron oxidation, to label the surface of viruses, living bacteria and cells. The functionalization was performed electrochemically and in situ by applying an electric potential to aqueous buffered solutions of tagged NML containing the viruses, bacteria or cells. The broad applicability of the click-electrochemistry method was explored on recombinant adeno-associated viruses (rAAV2), Escherichia coli (Gram-) and Staphyloccocus epidermidis (Gram + ) bacterial strains, and HEK293 and HeLa eukaryotic cell lines. Surface electro-conjugation was achieved in minutes to yield functionalized rAAV2 that conserved both structural integrity and infectivity properties, and living bacteria and cell lines that were still alive and able to divide.

摘要

目前,微生物表面的直接共价连接方法仍鲜有报道,且大多基于细菌和细胞的代谢工程功能化。虽然这些方法有效,但如果有一种更快的方法可以避免生物结合步骤,将具有很强的互补性。在这里,我们使用 N-甲基鲁米诺(NML),一种在单电子氧化后完全对酪氨酸具有选择性的蛋白锚定基团,对病毒、活菌和细胞的表面进行标记。通过对含有标记 NML 的水缓冲溶液施加电势,可以在原位电化学条件下对病毒、细菌或细胞进行功能化。该点击电化学方法的广泛适用性在重组腺相关病毒(rAAV2)、大肠杆菌(革兰氏阴性菌)和表皮葡萄球菌(革兰氏阳性菌)菌株以及 HEK293 和 HeLa 真核细胞系上进行了探索。在数分钟内实现了表面电共轭,得到了保持结构完整性和感染性的功能性 rAAV2,以及仍然存活并能够分裂的活菌和细胞系。

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