Self-Induced Back-Action Actuated Nanopore Electrophoresis (SANE) Sensor for Label-Free Detection of Cancer Immunotherapy-Relevant Antibody-Ligand Interactions.
Self-Induced Back-Action Actuated Nanopore Electrophoresis (SANE) Sensor for Label-Free Detection of Cancer Immunotherapy-Relevant Antibody-Ligand Interactions.
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DOI：
10.1007/978-1-0716-1811-0_20
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发表时间：
2022
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我们通过集成固态纳米孔和双纳米孔纳米孔径制造了一种新型单分子纳米传感器。该纳米传感器采用自诱导反作用 (SIBA) 进行光学捕获，并启用 SIBA 驱动纳米孔电泳 (SANE) 以同时采集分子相互作用的双峰光学和电学特征。这项工作描述了如何制造和使用 SANE 传感器来量化抗体-配体相互作用。我们描述了如何分析双峰光电数据，以与单独的电测量相比，改进抗体和配体与结合复合物的区分。描述了针对 T 细胞受体样抗体 (TCRmAb) 的特异性相互作用检测的示例结果，该抗体被设计用于靶向肽呈递主要组织相容性复合体 (pMHC) 配体，代表癌细胞表面上呈递的靶配体模型。我们还描述了如何分析双峰光电数据以区分抗体和配体之间的特异性和非特异性相互作用。显示了针对相同 pMHC（作为对照）的与癌症无关的 TCRMAb 的非特异性相互作用的示例结果。这些示例结果证明了 SANE 传感器作为癌症免疫治疗中配体靶点的潜在筛选工具的实用性，尽管我们相信其潜在用途要广泛得多。
We fabricated a novel single molecule nanosensor by integrating a Solid-State Nanopore and a Double Nanohole nanoaperture. The nanosensor employs Self-Induced Back-Action (SIBA) for optical trapping and enables SIBA-Actuated Nanopore Electrophoresis (SANE) for concurrent acquisition of bimodal optical and electrical signatures of molecular interactions. This work describes how to fabricate and use the SANE sensor to quantify antibody-ligand interactions. We describe how to analyze the bimodal optical-electrical data to improve upon the discrimination of antibody and ligand versus bound complex compared to electrical measurements alone. Example results for specific interaction detection are described for T-cell receptor-like antibodies (TCRmAbs) engineered to target peptide-presenting Major Histocompatibility Complex (pMHC) ligands, representing a model of target ligands presented on the surface of cancer cells. We also describe how to analyze the bimodal optical-electrical data to discriminate between specific and non-specific interactions between antibodies and ligands. Example results for non-specific interactions are shown for cancer-irrelevant TCRmAbs targeting the same pMHCs, as a control. These example results demonstrate the utility of the SANE sensor as a potential screening tool for ligand targets in cancer immunotherapy, though we believe that its potential uses are much broader.