Confined Gel-Electromembrane Extraction of Oligonucleotides

Area of Science:

• Analytical Chemistry
• Biotechnology
• Separation Science

Background:

• Gel-electromembrane extraction (G-EME) is a green analytical technique but is hindered by electroendosmosis (EEO) flow.
• EEO flow negatively impacts the efficiency and reproducibility of G-EME.

Purpose of the Study:

• To introduce confined gel-electromembrane extraction (CG-EME) to mitigate EEO flow.
• To develop a 3D-printed device for implementing CG-EME.
• To demonstrate CG-EME's effectiveness in extracting nucleic acids.

Main Methods:

• A novel 3D-printed modular device was designed to limit sample compartment volume, thus confining EEO flow.
• A mesh structure was incorporated to create thin, stable gel films, enhancing electromigration.
• Polar oligonucleotides were extracted, followed by optimization of voltage and gel parameters.

Main Results:

• The CG-EME method successfully extracted 62% of oligonucleotides in 15 minutes at 50 V using a 3% agarose gel film.
• The technique was validated by recovering DNA primers and isolating miRNA-181b from clinical samples.
• Upregulation of miRNA-181b in schizophrenia patients was detected using CG-EME coupled with qPCR.

Conclusions:

• CG-EME effectively suppresses EEO flow, enhancing electromembrane extraction performance.
• The developed 3D-printed device enables efficient nucleic acid isolation for biomarker discovery.
• CG-EME shows significant potential for clinical applications, expanding the utility of EME technology.