Dendron-Modified Polystyrene Microtiter Plate: Surface Characterization with Picoforce AFM and Influence of Spacing between Immobilized Amyloid Beta Proteins
Dendron-Modified Polystyrene Microtiter Plate: Surface Characterization with Picoforce AFM and Influence of Spacing between Immobilized Amyloid Beta Proteins
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DOI：
10.1021/la801872r
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发表时间：
2008-12-16
期刊：
影响因子：
3.9
通讯作者：
Park, Joon Won
中科院分区：
文献类型：
--
作者：
Roy, Dhruvajyoti;Kwak, Ju-Won;Park, Joon Won
将聚苯乙烯微滴定板涂覆一层锥形树枝状分子层，作为一种在固定生物分子之间提供适当间距的手段。在涂层制备过程中，通过等离子体将二(乙二醇基)乙烯基醚接枝到微滴定板表面，然后第二代树枝状分子(9-酸)或第三代树枝状分子(27-酸)自组装。接触角分析表明，等离子体接枝后亲水性显著增加，而树枝状固定化后亲水性恢复/降低。为了用力原子力显微镜(AFM)进行分析，将寡核苷酸固定在AFM尖端和平板上。观察到DNA-DNA相互作用的所有斑点，这表明树突的涂层在整个表面是均匀的。通过进行酶联免疫吸附试验(ELISA)，与其他常规平板(如未经处理的聚苯乙烯或顺丁烯二酸酐活化平板)相比，观察到对不同片段的淀粉样β蛋白(Aβ)的增强检测，从而检验了树枝状修饰的微滴定板上间距的生物分子分析的有效性。证实了生物分子间的间隔性对微滴定板的积极影响。
A polystyrene microtiter plate was,coated with a molecular layer of a cone-shaped dendron as a means of providing proper spacing between immobilized biomolecules. For the coating preparation, di(ethylene glycol) vinyl ether was grafted onto the surface of the microtiter plate by a plasma process followed by self-assembly of a second-generation dendron (9-acid) or a third-generation dendron (27-acid). Contact angle analysis revealed a pronounced increase in the hydrophilicity upon plasma grafting, while the hydrophilicity reverted/decreased after dendron immobilization. For analysis by force-based atomic force microscopy (AFM), oligonucleotides were immobilized onto the AFM tip and the plate. The DNA-DNA interaction was observed at all spots examined, which implied that coating of the dendrons was uniform over the entire surface. The effectiveness for biomolecular assays of the spacing on dendron-modified microtiter plates was examined by carrying out an enzyme-linked immunosorbent assay (ELISA), where enhanced detection of different fragments of amyloid beta protein (A beta) was observed when compared with other conventional plates, such as untreated polystyrene or maleic anhydride activated plates. The positive influence of the mesospacing between biomolecules on the microtiter plates for this assay was confirmed.