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Engineering Aptamer with Enhanced Affinity by Triple Helix-Based Terminal Fixation.

Lianhui Zhao1, Xiaoyan Qi1, Xiaochen Yan1

  • 1College of Food Science and Engineering , Ocean University of China , Qingdao , Shandong 266003 , China.

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|October 15, 2019
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Researchers engineered more stable aptamers by fixing their ends with a triple helix structure. This improved aptamer affinity 10-fold and enabled a highly sensitive electrochemical aptasensor for lysozyme detection in wine.

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Area of Science:

  • Biotechnology
  • Molecular Engineering
  • Biosensor Development

Background:

  • Aptamer flexibility often hinders stable structure formation and optimal binding.
  • Engineering stabilized aptamer structures is crucial for enhanced affinity and sensor performance.

Purpose of the Study:

  • To develop a postselection strategy for stabilizing aptamer structures.
  • To enhance the affinity and performance of an anti-lysozyme aptamer.
  • To create a sensitive electrochemical aptasensor for lysozyme detection in complex matrices.

Main Methods:

  • Stabilizing aptamer termini using a length-optimized triple helix structure.
  • Engineering an anti-lysozyme aptamer via this postselection strategy.
  • Developing an electrochemical aptasensor utilizing a DNA tetrahedron spacer.

Main Results:

  • The engineered aptamer exhibited a nearly 10-fold increase in affinity for lysozyme.
  • The electrochemical aptasensor showed a 180-fold lower limit of detection compared to controls.
  • The aptasensor demonstrated high sensitivity and selectivity for lysozyme in real red wine samples.

Conclusions:

  • Termini fixation via triple helix structures effectively stabilizes aptamers and enhances affinity.
  • The engineered aptamer and aptasensor show significant potential for detecting analytes in complex sample matrices.
  • This strategy offers a promising approach for aptamer engineering and biosensor applications.