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High-Throughput Strategy for Enhancing Aptamer Performance across Different Environmental Conditions.

Leighton Wan1, Alex Yoshikawa2, Michael Eisenstein2,3

  • 1Department of Bioengineering, Stanford University, Stanford, California 94305, United States.

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|June 14, 2023
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Summary

Researchers developed a high-throughput screening method to adapt aptamers for new environments. This technique significantly improved a glucose aptamer's binding affinity in physiological conditions after one screening round.

Keywords:
aptamersbiosensorshigh-throughput screeningphysiologicalstructure-switching

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biochemistry

Background:

  • Aptamers selected in specific conditions often lose affinity in different environments, limiting their biomedical use.
  • Biomedical applications require aptamers to function in complex biological samples like blood or urine.

Purpose of the Study:

  • To develop a high-throughput screening procedure for adapting existing aptamers to new environmental conditions.
  • To enhance aptamer affinity for specific assay conditions, such as physiological environments.

Main Methods:

  • Utilized a modified DNA sequencer for high-throughput screening of up to 10^7 aptamer mutants.
  • Screened all single- and double-substitution mutants of a glucose aptamer selected in high-ionic strength buffer.

Main Results:

  • Identified aptamer mutants with approximately four-fold increased affinity in physiological conditions after one screening round.
  • Observed modest improvements from single-base substitutions, but significant binding enhancements from double mutants, indicating cooperative effects.

Conclusions:

  • The high-throughput screening approach effectively adapts aptamers for altered environmental conditions.
  • This method is generalizable to various aptamers and conditions, offering a powerful tool for aptamer engineering in diverse applications.