Sensors 2011, 11(10), 9426-9441; doi:10.3390/s111009426
Article

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

1 Dipartimento di Scienze Farmaceutiche, University of Padova, via F. Marzolo 5, I-35131 Padova, Italy 2 Associazione CIVEN, via delle industrie 9, I-30175 Venezia-Marghera, Italy
* Author to whom correspondence should be addressed.
Received: 29 July 2011; in revised form: 27 September 2011 / Accepted: 28 September 2011 / Published: 3 October 2011
(This article belongs to the Special Issue Aptamer-Based Sensors)
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Abstract: 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.
Keywords: aptamer; thrombin; sandwich aptamer microarray

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MDPI and ACS Style

Sosic, A.; Meneghello, A.; Cretaio, E.; Gatto, B. Human Thrombin Detection Through a Sandwich Aptamer Microarray: Interaction Analysis in Solution and in Solid Phase. Sensors 2011, 11, 9426-9441.

AMA Style

Sosic A, Meneghello A, Cretaio E, Gatto B. Human Thrombin Detection Through a Sandwich Aptamer Microarray: Interaction Analysis in Solution and in Solid Phase. Sensors. 2011; 11(10):9426-9441.

Chicago/Turabian Style

Sosic, Alice; Meneghello, Anna; Cretaio, Erica; Gatto, Barbara. 2011. "Human Thrombin Detection Through a Sandwich Aptamer Microarray: Interaction Analysis in Solution and in Solid Phase." Sensors 11, no. 10: 9426-9441.

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