期刊
SENSORS
卷 11, 期 10, 页码 9426-9441出版社
MDPI
DOI: 10.3390/s111009426
关键词
aptamer; thrombin; sandwich aptamer microarray
资金
- FU-PAT (Provincia Autonoma di Trento) Project NAOMI
- Progetti di Ricerca di Ateneo, University of Padova
We have developed an aptamer-based microarray for human thrombin detection exploiting two non-overlapping DNA thrombin aptamers recognizing different exosites of the target protein. The 15-mer aptamer (TBA1) binds the fibrinogen-binding site, whereas the 29-mer aptamer (TBA2) binds the heparin binding domain. Extensive analysis on the complex formation between human thrombin and modified aptamers was performed by Electrophoresis Mobility Shift Assay (EMSA), in order to verify in solution whether the chemical modifications introduced would affect aptamers/protein recognition. The validated system was then applied to the aptamer microarray, using the solid phase system devised by the solution studies. Finally, the best procedure for Sandwich Aptamer Microarray (SAM) and the specificity of the sandwich formation for the developed aptasensor for human thrombin were optimized.
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