Multiplexed TCR and BCR mRNA Amplification for Long-Read Sequencing

Analyzing T-cell receptors (TCRs) and B-cell receptors (BCRs) is important for understanding immune responses, disease progression, and therapeutic outcomes. However, profiling multiple immune receptor types across many samples can require complex workflows and make it difficult to efficiently track individual samples during sequencing. Capturing diverse receptor chains while preserving sample identity is particularly important for comprehensive immune profiling. There is a need for a streamlined approach that prepares and distinguishes multiple immune receptor sequences for long-read analysis.

 

Researchers at Arizona State University have developed a streamlined workflow for multiplexed amplification and long-read sequencing of T-cell and B-cell receptor transcripts across multiple samples. By reverse transcribing, amplifying and barcode-labeling TCR and BCR mRNA, the resulting cDNA can be downstream long-read sequenced. This approach enables multiple samples and immune receptor types to be efficiently processed and distinguished during downstream sequencing. Receptor-specific primers support analysis across a range of immune receptor chains, while barcodes preserve sample identity during multiplexed processing. The protocol has detected specific TCRs at abundances below 5%.

 

This technology provides a streamlined workflow which permits rapid, portable TCR and BCR sequencing for repertoire analysis and antigen specific discovery.