Influence of Precise Polymer Conjugation on Aptamer-Target Binding

Area of Science:

• Bioconjugation Chemistry
• Nucleic Acid Aptamer Technology
• Biomolecular Engineering

Background:

• The ATP aptamer is a DNA sequence that binds adenosine triphosphate (ATP).
• Polymer bioconjugation is a technique used to modify biomolecules with synthetic polymers.
• Understanding how polymer conjugation affects aptamer binding is crucial for developing aptamer-based applications.

Purpose of the Study:

• To investigate the impact of polymer bioconjugation on the binding properties of the ATP aptamer.
• To synthesize and characterize novel polymer-aptamer bio-hybrids with varying architectures.
• To evaluate the functionality of the ATP aptamer after conjugation with synthetic poly(phosphodiester) chains.

Main Methods:

• Synthesis of ten different polymer-aptamer bio-hybrids using automated solid-phase phosphoramidite chemistry.
• Characterization of bio-hybrids using electrospray mass spectrometry, ion-exchange HPLC, and polyacrylamide gel electrophoresis.
• Binding studies using bio-layer interferometry (BLI) and surface plasmon resonance (SPR) to assess ATP binding affinity.

Main Results:

• Successful synthesis and characterization of ten polymer-aptamer bio-hybrids with diblock and triblock architectures.
• All synthesized bio-hybrid macromolecules containing the ATP aptamer sequence demonstrated ATP binding.
• Polymer conjugation did not compromise the ATP aptamer's functionality, regardless of its position within the bio-hybrid structure.

Conclusions:

• Polymer bioconjugation is a viable strategy for modifying ATP aptamers without compromising their target-binding capabilities.
• The synthesized polymer-aptamer bio-hybrids maintain aptamer functionality, opening possibilities for advanced aptamer-based sensors and therapeutics.
• This study provides valuable insights into the design and application of aptamer-polymer conjugates in various fields.