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Development and Optimization of a Thrombin Sandwich Aptamer Microarray.

Anna Meneghello1, Alice Sosic2, Agnese Antognoli3

  • 1Associazione CIVEN, Via delle Industrie 9, I-30175 Venezia-Marghera, Italy. meneghello@civen.org.

Microarrays (Basel, Switzerland)
|September 9, 2016
PubMed
Summary

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This study optimized an aptamer microarray for detecting human thrombin. The assay shows high specificity and sensitivity, achieving sub-nanomolar detection limits for thrombin in biological samples.

Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry

Background:

  • Thrombin is a key protein in blood coagulation.
  • Accurate detection of thrombin is crucial for diagnosing and monitoring various medical conditions.
  • Existing detection methods may lack sensitivity, specificity, or cost-effectiveness.

Purpose of the Study:

  • To optimize a sandwich aptamer microarray for sensitive and specific detection of human thrombin.
  • To evaluate the performance of the aptamer microarray in biological matrices like serum.
  • To establish a low-cost, rapid assay for thrombin detection.

Main Methods:

  • Development and optimization of a sandwich microarray using two distinct aptamers (TBA1 and TBA2) for human thrombin.
  • Utilizing TBA1 as the capture layer and TBA2 as the detection layer.
Keywords:
aptamerbioassayfluorescencemultiplexing microarraythrombin

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  • Assessing sensitivity via direct fluorescent analysis (Cy5-labeled TBA2) and indirect detection (TBA2-biotin/streptavidin).

    • Main Results:

    • The optimized aptamer microarray achieved high specificity for human thrombin.
    • Sub-nanomolar limits of detection (LODs) were obtained under various detection conditions.
    • The assay demonstrated reliable performance in the presence of human serum.

    Conclusions:

    • The developed aptamer microarray is a sensitive, specific, and cost-effective method for human thrombin detection.
    • The two-site binding assay format ensures high specificity compatible with routine analysis.
    • This technology holds promise for clinical diagnostics and biological sample analysis.