Dynamic selection of high-affinity aptamers using a magnetically activated continuous deflection microfluidic chip.

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Aptamer discovery is crucial for molecular diagnostics and therapeutics.
• Traditional aptamer selection methods can be time-consuming and inefficient.
• Developing faster, more effective aptamer selection techniques is essential.

Purpose of the Study:

• To design and validate a magnetically activated continuous deflection (MACD) chip for accelerated aptamer discovery.
• To demonstrate the efficacy of dynamic selection in enriching high-affinity aptamers.
• To obtain a high-affinity and high-specificity aptamer for Candida albicans.

Main Methods:

• A MACD chip was engineered to facilitate continuous flow dynamic selection.
• Aptamers were selected against Candida albicans using the MACD chip over six rounds.
• Selection stringency was modulated by adjusting target concentration.
• Aptamer binding affinity was quantified using the dissociation constant (Kd).

Main Results:

• The MACD chip enabled consecutive binding and separation in a continuous flow system.
• Dynamic selection efficiently eluted low-affinity sequences and enriched high-affinity aptamers.
• A C.al3 aptamer with high affinity and specificity for Candida albicans was successfully isolated.
• The selected aptamer exhibited a dissociation constant (Kd) of 7.9 nM.

Conclusions:

• Dynamic selection utilizing a MACD chip is a highly effective strategy for rapid aptamer discovery.
• The developed MACD chip technology accelerates the identification of high-affinity aptamers.
• The isolated C.al3 aptamer shows promise for applications targeting Candida albicans.