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Click-Particle Display for Base-Modified Aptamer Discovery.

Chelsea K L Gordon, Diana Wu, Anusha Pusuluri

  • 1Department of Chemistry , Boston College , Chestnut Hill , Massachusetts 02467 , United States.

ACS Chemical Biology
|September 19, 2019
PubMed
Summary
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We developed a click-chemistry and fluorescence-activated cell sorting (FACS) method to rapidly generate high-affinity, specific base-modified aptamers. This approach enables efficient screening of millions of aptamers per hour for diverse applications.

Area of Science:

  • Biotechnology
  • Chemical Biology
  • Molecular Biology

Background:

  • Base-modified aptamers offer superior affinity and specificity compared to natural DNA/RNA aptamers.
  • Traditional methods for creating modified aptamers are resource-intensive and require specialized expertise.

Purpose of the Study:

  • To develop a generalized and accelerated method for generating base-modified aptamers.
  • To enable high-throughput screening of aptamers for both affinity and specificity.

Main Methods:

  • Combined click-chemistry with fluorescence-activated cell sorting (FACS) for aptamer screening.
  • Developed a "click-particle display (PD)" strategy for rapid, high-throughput aptamer generation and selection.
  • Utilized FACS to measure quantitative on- and off-target binding of individual aptamers at a throughput of ~10^7 per hour.

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Main Results:

  • Generated a boronic acid-modified aptamer with ~1 μM affinity for epinephrine, a novel aptamer target.
  • Developed a mannose-modified aptamer with nanomolar affinity for concanavalin A (Con A).
  • Demonstrated that chemical modifications are essential for aptamer binding and that the Con A aptamer exhibits high specificity and potent biological activity, inhibiting Con A-mediated hemagglutination.

Conclusions:

  • The click-PD strategy significantly accelerates and simplifies the generation of base-modified aptamers.
  • This method allows for simultaneous optimization of affinity and specificity, leading to highly effective aptamers.
  • The click-PD approach is readily adoptable due to the widespread availability of FACS instrumentation.