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Integrated Microfluidic Isolation of Aptamers Using Electrophoretic Oligonucleotide Manipulation
Jinho Kim1, Timothy R Olsen1, Jing Zhu1
1Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
Scientific Reports
|May 25, 2016
Summary
We developed a microfluidic system for rapid DNA aptamer isolation using systematic evolution of ligands by exponential enrichment (SELEX). This integrated approach significantly reduces time and material, enabling versatile aptamer development for diverse targets.
Area of Science:
- Biotechnology
- Molecular Biology
- Analytical Chemistry
Background:
- Systematic evolution of ligands by exponential enrichment (SELEX) is crucial for aptamer discovery.
- Traditional SELEX methods are time-consuming and resource-intensive.
- Integrating SELEX processes in microfluidics offers potential for improved efficiency.
Purpose of the Study:
- To present a microfluidic approach for integrated DNA aptamer isolation via SELEX.
- To demonstrate a microbead-based and electrophoretic DNA manipulation scheme for SELEX.
- To enable highly efficient aptamer isolation with reduced time and material consumption.
Main Methods:
- Microfluidic system integrating microbead-based affinity selection and amplification.
- Electrophoretic DNA manipulation to couple processes in different buffers.
- Demonstrated on isolating aptamers against surface-immobilized immunoglobulin E and solution-phase bisboronic acid.
Main Results:
- Achieved full microfluidic integration of SELEX.
- Successfully isolated aptamer candidates in three rounds within approximately 10 hours.
- Demonstrated broad target applicability for both surface-immobilized and solution-borne targets.
Conclusions:
- The microfluidic SELEX approach enables highly efficient aptamer isolation.
- This method drastically reduces SELEX process time and biological material requirements.
- The approach facilitates the development of aptamers as versatile affinity reagents for various targets.
