4. Single-Cell AIR Profiling

Single-cell immune receptor profiling is a revolutionary approach that allows investigators to combine clonotype repertoire identification with paired-chain information and the phenotype of cells (e.g., cell subtype). Single-cell immune receptor profiling can be performed using a medium-throughput approach (1,000-5,000 cells) using microwell arrays or droplet microfluidics technologies. However, these assays are more complicated to run and require expensive reagents and limited sequencing throughput when compared to bulk immune receptor profiling methods.

Cellecta has developed a low-throughput, single-cell immune profiling strategy using sorted cells in 96-well plates. The plates are pre-aliquoted with T-cell receptor (TCR) α/β and TCR γ/δ primers along with 30 crucial T-cell markers. We perform multiplex RT-PCR amplification and sequencing of the CDR3 regions. The resulting data provides the abundant clonotype sequences along with the chain pairing information for these TCR α/β and γ/δ chains, along with the T-cell subtype information using gene-expression profiles. By analyzing the TCR gene rearrangement at the single-cell level, researchers can better understand T-cell development, proliferation, and clonality, which are crucial for studying diseases such as cancer, immunodeficiency, and autoimmunity. Furthermore, single-cell TCR sequencing provides a user-friendly tool for the development of potential T-cell-based cancer immunotherapies. The technology’s cost-effectiveness and ability to analyze clonotypes and immunophenotypes of cells in a single assay make it a valuable tool for the detection and characterization of antigen-activated TCRs.

This technology is currently available as a service. The kit will be available in Spring 2025.

To use this service, prepare and ship FACS-sorted single T- or B-cells plated individually in 96-well plates. Once received, the cells are processed to obtain receptor sequences and relevant expression markers. The resulting dataset includes clonotype abundances, chain pairing information, and expression profiles of key T- or B-cell markers. The final report provides a comprehensive overview of receptor diversity and highlights dominant or functionally significant clonotypes.