Human Thrombin Detection Through a Sandwich Aptamer Microarray: Interaction Analysis in Solution and in Solid Phase

Sensors

Alice Sosic²  Anna Meneghello¹  Erica Cretaio¹ 

[1] Associazione CIVEN, via delle industrie 9, I-30175 Venezia-Marghera, Italy; E-Mails:;Dipartimento di Scienze Farmaceutiche, University of Padova, via F. Marzolo 5, I-35131 Padova, Italy; E-Mail:

关键词: aptamer;    thrombin;    sandwich aptamer microarray;   

DOI  :  10.3390/s111009426

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2011 by the authors; licensee MDPI, Basel, Switzerland.