Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet
Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet
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DOI：
10.1016/j.ab.2008.03.035
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发表时间：
2008-06-15
影响因子：
2.9
通讯作者：
Pons, Jaurne
中科院分区：
文献类型：
--
作者：
Abdiche, Yasmina;Malashock, Dan;Pons, Jaurne
ForteBio的Octet光学生物传感器利用生物层干涉测量技术，使用一次性光纤生物传感器来检测和量化分子相互作用，该传感器可以在没有任何微流体的情况下从开放的振动微孔板中处理样品。我们招募了针对一组肽的单克隆抗体，将Octet直接与Biacore完善的3000平台和Bio-Rad最近推出的ProteOn XPR36阵列系统进行比较，后者使用表面等离子体共振（SPR）分别检测一种分析物在四个表面上的结合和六种分析物在六个表面上的结合。用沉淀法防止分析物重新结合配体包覆的八元体尖端，并使我们能够提取准确的动力学速率常数，因为它们与SPR测定的结果非常一致。虽然Octet的灵敏度不足以直接检测小分子的结合，但它可以通过溶液竞争实验间接获得它们的亲和力。我们在SPR仪器上进行了类似的实验来验证这些测量结果。Octet正在成为其他更复杂的生物传感器的多功能补充，ProteOn提供接近Biacore灵敏度的高质量数据，但采用更多的多路传输格式。我们的结果为评估上述传感器的性能提供了一个基准。(c) 2008爱思唯尔公司版权所有。
ForteBio's Octet optical biosensor harnesses biolayer interferometry to detect and quantify molecular interactions using disposable fiber-optic biosensors that address samples from an open shaking microplate without any microfluidics. We recruited a monoclonal antibody against a panel of peptides to compare the Octet directly with Biacore's well-established 3000 platform and Bio-Rad's recently launched ProteOn XPR36 array system, which use surface plasmon resonance (SPR) to detect the binding of one analyte over four surfaces and six analytes over six surfaces, respectively. A sink method was used to prevent analyte from rebinding the ligand-coated Octet tips and enabled us to extract accurate kinetic rate constants, as judged by their close agreement with those determined by SPR. Although the Octet is not sensitive enough to detect the binding of small molecules directly, it can access their affinities indirectly via solution competition experiments. We conducted similar experiments on the SPR instruments to validate these measurements. The Octet is emerging as a versatile complement to other more sophisticated biosensors, and the ProteOn provides high-quality data near the sensitivity of Biacore but in a more multiplexed format. Our results provide a benchmark for assessing the performance of the above-mentioned sensors. (c) 2008 Elsevier Inc. All rights reserved.