A radioisotope label-free alpha-bungarotoxin-binding assay using BIAcore sensor chip technology for real-time analysis.
A radioisotope label-free alpha-bungarotoxin-binding assay using BIAcore sensor chip technology for real-time analysis.
复制标题
使用 BIAcore 传感器芯片技术进行实时分析的无放射性同位素标记的 α-银环蛇毒素结合测定。
DOI：
10.1016/j.ab.2009.03.011
复制
发表时间：
2009
影响因子：
2.9
通讯作者：
Hawrot,Edward
中科院分区：
文献类型：
--
作者：
Paulo,JoaoA;Hawrot,Edward
α-银环蛇毒素（α-bgtx）结合蛋白，包括某些烟碱乙酰胆碱受体和乙酰胆碱结合蛋白（AChBPs），通常采用放射性同位素标记的α-bgtx结合试验进行表征。然而，这样的测定排除了在真实的时间内研究结合相互作用，并且受到与放射性同位素标记的试剂相关的不便的阻碍。我们使用基于表面等离子体共振的技术（BIAcore）来研究重组AChBP与直接固定α-bgtx的CM-5传感器芯片表面的结合。我们通过使用传统的125 I标记的α-bgtx结合试验比较相同的生物样品来验证我们的BIAcore结果。如本文所述，α-bgtx传感器芯片可以进行详细的、实时的、无放射性同位素的相互作用研究，这可以极大地促进新型α-bgtx结合蛋白和复合物的表征。
α-Bungarotoxin (α-bgtx)-binding proteins, including certain nicotinic acetylcholine receptors and acetylcholine-binding proteins (AChBPs), are frequently characterized with radioisotope-labeled α-bgtx-binding assays. Such assays, however, preclude investigations of binding interactions in real time and are hampered by the inconveniences associated with radioisotope-labeled reagents. We used surface plasmon resonance-based technology (BIAcore) to investigate the binding of recombinant AChBP to CM-5 sensor chip surfaces with directly immobilized α-bgtx. We validated our BIAcore results by comparing the same biological samples using the traditional125I-labeled α-bgtx-binding assay. An α-bgtx sensor chip, as described here, enables detailed, real-time, radioisotope-free interaction studies that can greatly facilitate the characterization of novel α-bgtx-binding proteins and complexes.