Tunable Nanofiber Mesh Coatings for Improved Nanopores Sensing

描述（由申请人提供）：虽然固态纳米孔显示出作为单分子传感器用于生物医学应用的前景，但为了提高其分辨率和效率，分析物分子必须更频繁地被探测，并且必须在纳米孔感测体积中保持更长时间。该提案描述了一种新颖的、简单的和通用的方法，以通过减慢移位速度和经由纳米孔外部的相互作用增加捕获效率来改善纳米孔灵敏度和时间分辨率，同时仍然使纳米孔本身可用于进一步官能化。这将通过将化学可调的纳米聚合物网（NFM）直接应用于固态纳米孔芯片来实现，其中其纤维在从纳米孔外部感测期间，纳米孔（疏水的、阳离子的或阴离子的）可以与分析物分子相互作用，将它们引导到感测体积中并减缓它们通过感测体积的移动。（100-1000 nm直径），由聚（β-己内酯）（PCL）掺杂聚（甘油-co-？-己内酯）（PGC），通过静电纺丝沉积到表面上。该聚合物的PGC组分可以被改性以包括一系列侧基，其赋予NFM疏水性、阴离子性或阳离子性。纤维尺寸和网格密度可以通过调节静电纺丝参数来控制。因此，聚合物NFM参数的调整将完全独立于纳米孔表面本身的改变。通过简单选择适当的NFM，任何固态纳米孔应用都可以获得广泛的设计师易位性质。我们将首先构建具有不同电荷和网格特性的纳米孔-纳米网格（NP-NFM）混合设备，然后筛选这些设备以确定哪些组合适合用于纳米孔传感。将使用一系列ssDNA和dsDNA长度对候选器械进行测试，以确定每种类型的NFM如何影响易位和捕获。可以提高感测效率和分辨率而不需要对纳米孔进行内在修饰的外部化学可调涂层的应用将通过实现较慢的易位速度和增加的穿线概率来直接支持开发可行的基于纳米孔的测序和诊断平台的努力。

DESCRIPTION (provided by applicant): While solid-state nanopores show promise as single-molecule sensors for biomedical applications, in order to improve their resolution and efficiency, analyte molecules must be probed more frequently and must remain longer in the nanopore sensing volume. This proposal describes a novel, simple, and versatile method to improve nanopore sensitivity and temporal resolution by slowing translocation speed and increasing capture efficiency via interactions outside the nanopore, while still leaving the nanopore itself available for further functionalization. This will be achieved by applying a chemically tunable nanofiber polymeric mesh (NFM) directly to a solid-state nanopore chip, where its fibers (hydrophobic, cationic, or anionic) may interact with analyte molecules during sensing from outside the nanopore, guiding them into the sensing volume and slowing their movement through it. The NFM is composed of a network of fibers (100-1000 nm diameter) formed from poly(?-caprolactone) (PCL) doped with poly(glycerol-co-?-caprolactone) (PGC), deposited onto a surface by electrospinning. The PGC component of this polymer may be modified to include a range of side groups which confer hydrophobic, anionic, or cationic properties to the NFM. The fiber size and mesh density may be controlled by adjusting the electrospinning parameters. Thus, adjustment of the polymer NFM parameters will be completely independent of alterations to the nanopore surface itself. A broad range of designer translocation properties will be accessible for any solid-state nanopore application through simple selection of an appropriate NFM. We will first construct nanopore-nanofiber mesh (NP-NFM) hybrid devices with different charge and mesh characteristics, then screen these devices to determine which combinations are suitable for nanopore sensing. Candidate devices will be tested using a range of ssDNA and dsDNA lengths to determine how each type of NFM affects translocation and capture. The application of an external, chemically tunable coating that can improve sensing efficiency and resolution without requiring intrinsic modification of the nanopore will directly support efforts t develop viable nanopore-based sequencing and diagnostic platforms by enabling slower translocation speeds and increased threading probability.