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General Label-Free Fluorescent Aptamer Binding Assay Using Cationic Conjugated Polymers.

Pengbo Zhang1,2, Ke Qin1, Anand Lopez2

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Analytical Chemistry
|October 25, 2022
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Summary

A new cationic conjugated polymer (CCP) assay offers a general, cost-effective method for aptamer binding detection. This label-free fluorescence assay demonstrates high sensitivity and specificity for various targets.

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Area of Science:

  • Biochemistry
  • Materials Science
  • Analytical Chemistry

Background:

  • A need exists for general, cost-effective, and reliable aptamer binding assays.
  • Aptamer-based detection relies on conformational changes upon target binding, which can be transduced into a measurable signal.

Purpose of the Study:

  • To develop and validate a general, cost-effective, and reliable aptamer binding assay using cationic conjugated polymers (CCPs).
  • To investigate the feasibility of using CCP fluorescence changes to detect aptamer-target interactions.

Main Methods:

  • Utilized Poly(3-(3'-N,N,N-triethylamino-1'-propyloxy)-4-methyl-2,5-thiophene hydrochloride) (PMNT) as a model CCP.
  • Optimized buffer conditions to 100 mM NaCl and 10 mM MgCl2 for aptamer binding.
  • Characterized four aptamers (K+, adenosine, cortisol, caffeine) using fluorescence spectroscopy, monitoring changes in peak intensity or ratios.

Main Results:

  • The CCP-based assay demonstrated sensitive and specific detection of aptamer-target binding.
  • Quantification strategies varied based on aptamer structure (peak intensity drop or fluorescence ratio).
  • The assay showed comparable or superior sensitivity to SYBR Green I dye-based methods.

Conclusions:

  • The developed CCP-based assay is a practicable, general method for aptamer binding evaluation.
  • The assay is label-free, quick, cost-effective, and suitable for target detection as a biosensor.