Microfluidic Print-to-Synthesis Platform for Efficient Preparation and Screening of Combinatorial Peptide Microarrays

Area of Science:

• Biotechnology
• Chemical Engineering
• Drug Discovery

Background:

• Combinatorial synthesis enables rapid generation of diverse molecular libraries.
• Peptide microarrays are valuable tools for screening molecular interactions.
• Existing methods for peptide synthesis and screening can be time-consuming and reagent-intensive.

Purpose of the Study:

• To introduce a novel Microfluidic Print-to-Synthesis (MPS) platform for automated peptide microarray synthesis.
• To demonstrate the MPS platform's capability for high-throughput synthesis of unique peptide sequences.
• To validate the MPS platform using a cell-based assay for screening peptide interactions.

Main Methods:

• Utilized standard Fmoc chemistry on a polyethylene glycol (PEG)-functionalized microdisc array.
• Developed a microfluidic cartridge for dispensing building blocks and coupling reagents.
• Synthesized a focused tetrapeptide library of 2560 unique sequences.
• Screened the peptide microarray using live human T lymphocyte Jurkat cells to identify interactions with α4β1 integrin.

Main Results:

• Successfully synthesized peptide microarrays with 200 μm spot size and 1 mm center-to-center distance, minimizing reagent consumption.
• Demonstrated the scalability and multiplexibility of the MPS platform.
• Identified specific peptide sequences interacting with α4β1 integrin on Jurkat cells.
• Showcased the spatial addressability of the MPS array for direct identification of active compounds.

Conclusions:

• The MPS platform offers a novel, automated, and high-throughput solution for custom peptide microarray synthesis.
• MPS facilitates rapid screening and optimization of lead compounds through efficient biochemical or cell-based assays.
• This technology significantly advances the field of combinatorial chemistry and drug discovery.