cloneXplorer: A high-throughput clone discovery platform based on conical microwell arrays.

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Generating antigen-specific T cell populations is crucial for understanding immune recognition but traditionally involves laborious methods like limiting dilution or cloning.
• The need for efficient and high-throughput methods to isolate and characterize rare antigen-specific T cells is significant in immunological research.

Purpose of the Study:

• To develop and validate a streamlined approach for generating antigen-specific T cell clones directly from peripheral blood.
• To showcase the capabilities of the cloneXplorer live-cell analysis and clone isolation platform for identifying and recovering functionally defined T cells.

Main Methods:

• Utilized the cloneXplorer platform, a conical microwell array system, for live-cell analysis and clone isolation.
• Monitored cell proliferation, cytokine secretion (Interferon gamma), and surface markers in single-cell co-cultures.
• Employed fluorescent sandwich assays for immune activation monitoring and tetramer staining for specificity verification.

Main Results:

• Demonstrated efficient generation of monoclonal cell populations from cell lines and human T cells.
• Successfully identified antigen-specificity at single-cell resolution using engineered Jurkat cells and antigen-presenting cells with a peptide library.
• Showcased isolation and expansion of antigen-specific T cells from human peripheral blood based on Interferon gamma secretion and CD8 marker expression.

Conclusions:

• The cloneXplorer platform offers a powerful and efficient solution for generating antigen-specific T cell clones.
• This technology has broad applications in cell line development, immune receptor screening, and validation of immune responses.