High-throughput microfluidic chip with silica gel-C18 channels for cyclotide separation

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Conventional chromatography methods for biomolecule separation are costly and inefficient.
• Cyclotides, cyclic peptides from plants, exhibit diverse biological activities, including antimicrobial and anticancer properties, making them attractive for drug development.
• Efficient separation of cyclotides is crucial for their application in drug discovery.

Purpose of the Study:

• To develop and demonstrate a novel microfluidic chip-based separation method for cyclotides.
• To mimic high-performance liquid chromatography (HPLC) separation on a microfluidic platform.
• To assess the efficiency and applicability of microfluidic separation for cyclotide purification.

Main Methods:

• A microfluidic chip was prepared by packing synthesized silica gel-C18 into a microchannel, creating an HPLC C18 column mimic.
• Cyclotide extract from Viola ignobilis, containing specific cyclotides, was obtained using low voltage electric field extraction.
• The cyclotide extract was injected into the microchannel with distilled water as the mobile phase, and samples were collected at 2-minute intervals.

Main Results:

• Successful separation of individual cyclotides from the mixed extract within the microfluidic channel.
• Demonstration of distinct collection times for each separated cyclotide, indicating successful differentiation.
• Characterization of the cyclotide extract using HPLC and MALDI-TOF confirmed the presence of target cyclotides.

Conclusions:

• Microfluidic chip-based separation is a rapid, cost-effective, and simple method for separating cyclotides.
• This technology shows high impact and potential for widespread application in drug discovery research.
• The developed microfluidic device offers a viable alternative to conventional separation techniques for cyclotide purification.