Related Experiment Video
Updated: May 27, 2026
03:38
Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
Published on: October 6, 2022
Probing the limits of aptamer affinity with a microfluidic SELEX platform
Kareem M Ahmad1, Seung Soo Oh, Seon Kim
1Interdepartmental Program in Biomolecular Science and Engineering, University of California Santa Barbara, Santa Barbara, California, United States of America.
Plos One
|November 24, 2011
Summary
Microfluidic selection (M-SELEX) efficiently generates high-affinity aptamers for diagnostics and therapeutics. This method yielded superior DNA aptamers against PDGF-BB and thrombin, demonstrating potential for improved molecular recognition applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Biochemistry
Background:
- Nucleic acid aptamers offer advantages over antibodies, but their selection is time-consuming and often yields low-affinity molecules.
- Developing efficient and reproducible aptamer selection methods is crucial for molecular diagnostics and therapeutics.
Purpose of the Study:
- To develop a microfluidic selection process (M-SELEX) for generating high-affinity aptamers against diverse protein targets.
- To investigate the influence of protein charge on aptamer affinity.
Main Methods:
- A microfluidic selection process (M-SELEX) was employed to isolate DNA aptamers against three protein targets: PDGF-BB, thrombin, and ApoE.
- Aptamer selection was performed under varying pH conditions for PDGF-BB to study the effect of protein charge.
Main Results:
- Novel DNA aptamers were discovered for PDGF-BB (K(d) = 0.028 nM) and thrombin (K(d) = 0.33 nM) with affinities superior to previously reported aptamers.
- An aptamer for ApoE (K(d) = 3.1 nM) was also identified.
- An inverse correlation was observed between protein charge and aptamer dissociation constant (K(d)).
Conclusions:
- M-SELEX is an efficient method for generating high-performance aptamers against various protein targets.
- Protein charge significantly influences aptamer affinity, suggesting that charge modulation can optimize aptamer selection.
- The developed M-SELEX protocol holds promise for advancing aptamer-based diagnostics and therapeutics.
