Microfluidic-Enabled Production of DNA Barcoded APC Library (MEDAL) for High Throughput T Cell Epitope Screening
Xu Cui1,2, Yi Liu1,2, Tongjin Wu1,2
1Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore, Singapore, 117583, Singapore.
Small Methods
|May 15, 2025
Summary
This study introduces a microfluidic platform for rapid, high-throughput production of DNA-barcoded Antigen Presenting Cells (APCs). This innovation accelerates vaccine development by efficiently identifying peptide epitopes recognized by T cells.
Area of Science:
- Immunology
- Biotechnology
- Microfluidics
Background:
- Vaccine development requires screening peptide fragments for presentation on antigen-presenting cells (APCs).
- Current methods are slow and expensive due to chemical synthesis and individual cell pulsing.
- Efficiently generating and screening diverse peptide-APC interactions is crucial.
Purpose of the Study:
- To develop a high-throughput microfluidic platform for parallel production of DNA-barcoded APCs loaded with enzymatically synthesized peptides.
- To accelerate the identification of peptide epitopes for vaccine design.
- To enable rapid screening of up to 9000 different peptide-APC combinations.
Main Methods:
- Developed Microfluidic-Enabled production of DNA-barcoded APC Library (MEDAL) platform.
- Utilized microfluidic PCR-IVTT reactions to generate DNA-barcoded peptides within droplets.
- Injected APCs into droplets for parallel loading and barcode association.
- Employed next-generation sequencing for DNA barcode analysis.
Main Results:
- Successfully produced up to 9000 unique DNA-barcoded APCs within a 10-hour workflow.
- Identified peptide sequences binding to H-2Kb MHC class I molecules.
- Validated the platform by co-culturing T cells with MEDAL-prepared APC libraries to identify specific T cell epitopes.
Conclusions:
- MEDAL offers a rapid, cost-effective, and high-throughput solution for generating peptide-loaded APC libraries.
- This platform significantly advances vaccine development by streamlining epitope discovery.
- The technology enables efficient identification of T cell-recognized epitopes for targeted vaccine design.
