Electrochemical assay of concanavalin A-ovalbumin binding on magnetic beads.
Electrochemical assay of concanavalin A-ovalbumin binding on magnetic beads.
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DOI：
10.1039/c2an35667h
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发表时间：
2012-07
期刊：
影响因子：
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通讯作者：
K. Sugawara;Asako Yugami;T. Kadoya;H. Kuramitz;K. Hosaka
中科院分区：
文献类型：
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作者：
K. Sugawara;Asako Yugami;T. Kadoya;H. Kuramitz;K. Hosaka
为了监测磁珠上蛋白质-糖蛋白的相互作用，本研究建立了一种电化学分析法，测定了伴刀豆球蛋白A（ConA）和卵清蛋白（OVA）之间的结合。该系统是一个功能强大的模型，可用于评估细胞连接。将电活性柔红霉素（Daunomycin）与刀豆蛋白A（ConA）通过交联剂固定在6种不同大小的磁珠（直径1.0-8.9 μm）上。还使用类似的方法制备了六种尺寸的OVA珠（直径：1.0-8.9 μm）。使用ConA与柔红霉素的氧化峰评价结合，因为ConA识别具有α-甘露糖残基的OVA。当结合发生在珠的表面上时，由于电活性部分被OVA覆盖，峰值电流降低。当评价ConA/道诺霉素-OVA结合时，在存在OVA修饰的珠粒（直径：2.5 μm）的情况下，由ConA和道诺霉素修饰的珠粒（直径：8.9 μm）获得的峰电流的变化最大。相比之下，对于具有ConA/道诺霉素和OVA的最小珠（直径：1.0 μm）观察到颗粒团聚。结果表明，ConA-OVA结合依赖于珠的大小。因此，该方法可用于测量细胞表面的蛋白质-糖蛋白相互作用。
To monitor protein-glycoprotein interactions on magnetic beads, the present study developed an electrochemical assay of the binding between concanavalin A (ConA) and ovalbumin (OVA). The system was a powerful model that could be used to evaluate cell junctions. ConA with an electroactive daunomycin was immobilized on 6 different sizes of magnetic beads (diameter: 1.0-8.9 μm) through a cross-linking agent. Six sizes of OVA-beads (diameter: 1.0-8.9 μm) were also prepared using a similar method. The binding was evaluated using an oxidation peak of ConA with daunomycin because ConA recognized OVA with α-mannose residues. When binding took place on the beads' surface, the peak current was decreased due to the electroactive moieties being covered with OVA. When ConA/daunomycin-OVA binding was evaluated, the change of the peak current obtained by the beads (diameter: 8.9 μm) modified with ConA and daunomycin was the greatest in the presence of OVA-modified beads (diameter: 2.5 μm). In contrast, particle agglomeration was observed for the smallest beads (diameter: 1.0 μm) with ConA/daunomycin and OVA. The results suggested that ConA-OVA binding depended on the size of beads. Thus, this method could be applied to measure protein-glycoprotein interactions on the cell surface.