Two-dimensional membrane scaffold for the oriented immobilization of biosensing molecules
Two-dimensional membrane scaffold for the oriented immobilization of biosensing molecules
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DOI：
10.1016/j.bios.2019.111860
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发表时间：
2020-02-15
影响因子：
12.6
通讯作者：
Kuroda, Shun'ichi
中科院分区：
文献类型：
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作者：
Iijima, Masumi;Nakayama, Tsutomu;Kuroda, Shun'ichi
为了提高传感器的灵敏度和配体结合能力，需要精确控制传感器芯片上生物传感分子的取向和密度。我们先前开发了类似于30-nm生物纳米胶囊（ZZ-BNC），其由与串联形式的蛋白A衍生的IgG Fc结合Z结构域（ZZ-L（蛋白））融合的B病毒包膜L蛋白组成。这被用作稳健的纳米颗粒支架以增强IgG和Fc融合的传感分子（Fc融合受体）的敏感性和配体结合能力。然而，由于它们的刚性颗粒结构，ZZ-L蛋白的表面密度不能被优化用于生物传感器功能，并且无用的ZZ-L蛋白被卡在ZZ-BNC和传感器芯片之间。在这里，我们已经开发了一种平面脂质膜嵌入ZZ-L胶束（ZZ-L膜），它可以修改任何生物传感器芯片的表面与控制密度的ZZ-L蛋白。与ZZ-BNC相比，ZZ-L膜的IgG灵敏度和配体结合能力提高了约10倍。此外，固定化的IgG几乎可以化学计量地捕获它们各自的抗原，这表明ZZ-L膜在聚集和定向固定方面是Fc融合的传感分子的最理想的支架。
The orientation and density of biosensing molecules on sensor chip should be precisely controlled to improve sensitivity and ligand-binding capacity. We previously developed a similar to 30-nm bio-nanocapsule (ZZ-BNC), consisting of the hepatitis B virus envelope L protein fused with the tandem form of protein A-derived IgG Fc-binding Z domain (ZZ-L( protein). This is used as a robust nanoparticle scaffold to enhance the sensit)ivity and ligand-binding capacity of IgGs and Fc-fused sensing molecules (Fc-fused receptors). However, due to their rigid particle structure, the surface density of ZZ-L proteins could not be optimized for biosensor functions, and useless ZZ-L proteins become stuck between ZZ-BNC and the sensor chip. Here, we have developed a planar lipid membrane embedded with ZZ-L micelles (ZZ-L membrane), which could modify the surface of any biosensor chip with a controlled density of ZZ-L proteins. Compared with ZZ-BNC, the sensitivity and ligand-binding capacity of IgGs were enhanced about 10-fold with the ZZ-L membrane. Furthermore, the immobilized IgGs could capture their respective antigens almost stoichiometrically, indicating that ZZ-L membrane is the most ideal scaffold for Fc-fused sensing molecules in terms of both clustering and oriented immobilization.