Dick Jeffrey E, Hilterbrand Adam T, Boika Aliaksei, Upton Jason W, Bard Allen J
Department of Chemistry, Center for Electrochemistry, The University of Texas at Austin, Austin, TX 78712; and.
Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712.
Proc Natl Acad Sci U S A. 2015 Apr 28;112(17):5303-8. doi: 10.1073/pnas.1504294112. Epub 2015 Apr 13.
We report observations of stochastic collisions of murine cytomegalovirus (MCMV) on ultramicroelectrodes (UMEs), extending the observation of discrete collision events on UMEs to biologically relevant analytes. Adsorption of an antibody specific for a virion surface glycoprotein allowed differentiation of MCMV from MCMV bound by antibody from the collision frequency decrease and current magnitudes in the electrochemical collision experiments, which shows the efficacy of the method to size viral samples. To add selectivity to the technique, interactions between MCMV, a glycoprotein-specific primary antibody to MCMV, and polystyrene bead "anchors," which were functionalized with a secondary antibody specific to the Fc region of the primary antibody, were used to affect virus mobility. Bead aggregation was observed, and the extent of aggregation was measured using the electrochemical collision technique. Scanning electron microscopy and optical microscopy further supported aggregate shape and extent of aggregation with and without MCMV. This work extends the field of collisions to biologically relevant antigens and provides a novel foundation upon which qualitative sensor technology might be built for selective detection of viruses and other biologically relevant analytes.
我们报告了在超微电极（UME）上对鼠巨细胞病毒（MCMV）随机碰撞的观察结果，将UME上离散碰撞事件的观察扩展到了生物学相关分析物。通过一种针对病毒体表面糖蛋白的特异性抗体的吸附，在电化学碰撞实验中，根据碰撞频率的降低和电流大小，能够区分MCMV与被抗体结合的MCMV，这表明了该方法在对病毒样本进行大小测定方面的有效性。为了增加该技术的选择性，利用MCMV、一种针对MCMV的糖蛋白特异性一抗以及聚苯乙烯珠“锚定物”之间的相互作用来影响病毒的移动性，其中聚苯乙烯珠“锚定物”用针对一抗Fc区域的二抗进行了功能化修饰。观察到了珠子聚集现象，并使用电化学碰撞技术测量了聚集程度。扫描电子显微镜和光学显微镜进一步证实了有和没有MCMV时聚集体的形状和聚集程度。这项工作将碰撞领域扩展到了生物学相关抗原，并为构建定性传感器技术以选择性检测病毒和其他生物学相关分析物提供了一个新的基础。
