Division of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, OH, USA.
Oxford Nanopore Technologies Limited, Oxford Science Park, UK.
Nanomedicine. 2020 Apr;25:102170. doi: 10.1016/j.nano.2020.102170. Epub 2020 Feb 6.
The connector channel of bacteriophage phi29 DNA packaging motor has been inserted into the lipid bilayer membrane and has shown potential for the sensing of DNA, RNA, chemicals, peptides, and antibodies. Properties such as high solubility and large channel size have made phi29 channel an advantageous system for those applications; however, previously studied lipid membranes have short lifetimes, and they are frangible and unstable under voltages higher than 200 mV. Thus, the application of this lipid membrane platform for clinical applications is challenging. Here we report the insertion of the connector into the stable polymer membrane in MinION flow cell that contains 2048 wells for high-throughput sensing by the liposome-polymer fusion process. The successful insertion of phi29 connector was confirmed by a unique gating phenomenon. Peptide translocation through the inserted phi29 connector was also observed, revealing the potential of applying phi29 connector for high-throughput peptide sensing.
噬菌 Phi29 的 DNA 包装马达的连接通道已经插入脂质双层膜，并显示出对 DNA、RNA、化学物质、肽和抗体的传感潜力。高溶解度和大通道尺寸等特性使 Phi29 通道成为这些应用的优势系统；然而，之前研究的脂质膜寿命短，在高于 200 mV 的电压下易碎且不稳定。因此，该脂质膜平台在临床应用中的应用具有挑战性。在这里，我们报告了连接器插入 MinION 流动池中的稳定聚合物膜中，该流动池包含 2048 个孔，用于通过脂质体-聚合物融合过程进行高通量传感。通过独特的门控现象证实了 Phi29 连接器的成功插入。还观察到肽通过插入的 Phi29 连接器的易位，这表明应用 Phi29 连接器进行高通量肽传感的潜力。
