Adhesion mode atomic force microscopy study of dual component protein films

被引:22
作者
Agnihotri, A
Siedlecki, CA
机构
[1] Penn State Univ, Coll Med, Dept Surg, Inst Biomed Engn, Hershey, PA 17033 USA
[2] Penn State Univ, Coll Med, Dept Bioengn, Inst Biomed Engn, Hershey, PA 17033 USA
关键词
atomic force microscopy; adhesion mapping; fibrinogen; antibody-modified probes;
D O I
10.1016/j.ultramic.2004.10.006
中图分类号
TH742 [显微镜];
学科分类号
摘要
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.
引用
收藏
页码:257 / 268
页数:12
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