Measurement of small molecule binding kinetics on a protein microarray by plasmonic-based electrochemical impedance imaging.
Measurement of small molecule binding kinetics on a protein microarray by plasmonic-based electrochemical impedance imaging.
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DOI：
10.1021/ac5024556
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发表时间：
2014-10-07
影响因子：
7.4
通讯作者：
Tao, Nongjian
中科院分区：
文献类型：
--
作者：
Liang, Wenbin;Wang, Shaopeng;Festa, Fernanda;Wiktor, Peter;Wang, Wei;Magee, Mitchell;LaBaer, Joshua;Tao, Nongjian
我们报告的定量研究小分子结合动力学的蛋白质微阵列与基于等离子体的电化学阻抗显微镜（P-EIM）。P-EIM通过将表面电荷变化转换为表面等离子体共振（SPR）图像强度变化，以高空间分辨率光学测量电阻抗，并且信号不与分析物的质量成比例。使用P-EIM，我们测量了小分子药物（伊马替尼和SB 202190）与其靶蛋白（激酶β 1和p38-α）之间的结合动力学和亲和力。测得的亲和力值与间接竞争性结合测定法测得的报告值一致。我们还发现SB 202190与ABL 1具有弱结合，KD > 10 μM，这在文献中未见报道。此外，我们发现P-EIM不太容易发生非特异性结合，这是SPR中的一个长期问题。我们的研究结果表明，P-EIM是一种高通量测量小分子结合动力学和亲和力的新方法，这对于了解生物系统中的小分子和发现小分子药物至关重要。
We report on a quantitative study of small molecule binding kinetics on protein microarrays with plasmonic-based electrochemical impedance microscopy (P-EIM). P-EIM measures electrical impedance optically with high spatial resolution by converting a surface charge change to a surface plasmon resonance (SPR) image intensity change, and the signal is not scaled to the mass of the analyte. Using P-EIM, we measured binding kinetics and affinity between small molecule drugs (imatinib and SB202190) and their target proteins (kinases Abl1 and p38-α). The measured affinity values are consistent with reported values measured by an indirect competitive binding assay. We also found that SB202190 has weak bindings to ABL1 with KD > 10 μM, which is not reported in the literature. Furthermore, we found that P-EIM is less prone to nonspecific binding, a long-standing issue in SPR. Our results show that P-EIM is a novel method for high-throughput measurement of small molecule binding kinetics and affinity, which is critical to the understanding of small molecules in biological systems and discovery of small molecule drugs.
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影响因子：
3.8
作者：
Lesuffleur, Antoine;Im, Hyungsoon;Oh, Sang-Hyun
通讯作者：
Oh, Sang-Hyun
影响因子：
2.9
作者：
Papalia, Giuseppe A.;Leavitt, Stephanie;Myszka, David G.
通讯作者：
Myszka, David G.
影响因子：
2.9
作者：
Cannon, MJ;Papalia, GA;Myszka, DG
通讯作者：
Myszka, DG
影响因子：
11.4
作者：
Marina, A;Waldburger, CD;Hendrickson, WA
通讯作者：
Hendrickson, WA
影响因子：
11.2
作者：
Duensing, A;Joseph, NE;Fletcher, JA
通讯作者：
Fletcher, JA