Adhesion mode atomic force microscopy study of dual component protein films
Adhesion mode atomic force microscopy study of dual component protein films
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DOI：
10.1016/j.ultramic.2004.10.006
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发表时间：
2005-03-01
期刊：
影响因子：
2.2
通讯作者：
Siedlecki, CA
中科院分区：
文献类型：
--
作者：
Agnihotri, A;Siedlecki, CA
将原子力显微镜的分子识别成像扩展到云母上吸附的双组分蛋白质膜。AFM探针通过共价连接抗纤维蛋白原的多克隆抗体而功能化。利用原子力显微镜的附着力测绘模式生成地形图和附着力图像。功能化探针的有效性首先是通过在云母表面微接触印刷牛血清白蛋白然后回填纤维蛋白原形成的图案化双组分蛋白质膜上进行黏附映射来确定的。接下来，对随机分布的双组分蛋白单层进行粘附图，该单层是通过顺序吸附亚单层数量的纤维蛋白原，然后用牛血清白蛋白回填而产生的。粘附图被用来生成二值识别图像，其中基于统计得出的截止值来区分特定和非特定的相互作用。从识别图像获得的纤维蛋白原在完整的双蛋白单层上的表面覆盖率与用牛血清白蛋白回填之前获得的表面覆盖率相似。在用抗纤维蛋白原抗体封闭表面后，观察到的识别事件的数量减少了80%。这一结果表明，识别图像中的阳性事件确实是特异性的抗体-纤维蛋白原相互作用。(C)2004爱思唯尔B.V.保留所有权利。
Molecular recognition imaging by AFM was extended to dual component protein films adsorbed on mica. AFM probes were functionalized by covalently linking polyclonal antibodies against fibrinogen. Adhesion mapping mode of AFM was used to generate both topographic images and adhesion images. The efficacy of the functionalized probes was first established by performing adhesion mapping on patterned dual component protein films formed by microcontact printing bovine serum albumin on a mica surface and then backfilling with fibrinogen. Next, adhesion mapping was done on randomly distributed two-component protein monolayers generated by sequential adsorption of submonolayer amounts of fibrinogen followed by backfilling with bovine serum albumin. The adhesion maps were used to generate binary recognition images where the specific and non-specific interactions were differentiated based on a statistically derived cut-off value. The surface coverage of fibrinogen obtained from the recognition image over the complete dual protein monolayer was similar to that obtained prior to backfilling with bovine serum albumin. The number of recognition events that were observed decreased by >80% after blocking the surface with anti-fibrinogen antibodies. This result demonstrated that the positive events in the recognition image were indeed specific anti body-fibrinogen interactions. (C) 2004 Elsevier B.V. All rights reserved.