A label-free immunosensor array using single-chain antibody fragments
A label-free immunosensor array using single-chain antibody fragments
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DOI：
10.1073/pnas.0504917102
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发表时间：
2005-10-11
影响因子：
11.1
通讯作者：
Gerber, C
中科院分区：
文献类型：
--
作者：
Backmann, N;Zahnd, C;Gerber, C
我们报道了一种基于微电泳的免疫传感器，其在静态偏转模式下工作，其性能与表面等离子体共振相当，使用单链抗体（scFv）片段作为受体分子。作为模型系统，应用对两种不同抗原具有特异性的scFv片段。我们在每个scFv构建体的C末端引入半胱氨酸残基，以允许以定向方式共价连接到金涂覆的传感器界面。阵列的应用使得能够同时测量感测和参考杠杆的偏转。差异偏转信号显示特异性抗原结合，并且与溶液中的抗原浓度成比例。使用小的定向scFv片段作为受体分子，我们将微电泳的灵敏度提高到约1 nM。
We report a microcantilever-based immunosensor operated in static deflection mode with a performance comparable with surface plasmon resonance, using single-chain Fv (scFv) antibody fragments as receptor molecules. As a model system scFv fragments with specificity to two different antigens were applied. We introduced a cysteine residue at the C terminus of each scFv construct to allow covalent attachment to gold-coated sensor interfaces in directed orientation. Application of an array enabled simultaneous deflection measurements of sensing and reference cantilevers. The differential deflection signal revealed specific antigen binding and was proportional to the antigen concentration in solution. Using small, oriented scFv fragments as receptor molecules we increased the sensitivity of microcantilevers to approximate to 1 nM.