DriverMap AIR is the only assay technology on the market that profiles the full-length receptor region (exclusively RNA) or only the CDR3 repertoire of all T-cell receptor (TCR) - TRA, TRB, TRG and TRD - and B-cell receptor (BCR) - IGH, IGK and IKL - chains in an single-tube, single-day assay using multiplex PCR-NGS technology without the need of any additional specialized equipment. Cellecta offers AIR assays to profile human or mouse RNA, DNA, or both. The simultaneous profiling of DNA and RNA enables the identification of antigen-activated clonotypes to provide even greater insight into the immune response.
DriverMap Multiplex PCR technology uses gene-specific primers which significantly reduce the level of non-specific binding and primer-dimer amplification products and are designed to target only expressed TCR/BCR mRNA isoforms. Unique Molecular Identifiers (UMIs) facilitate accurate quantitation of the copy number of cDNA molecules in amplification steps, as well as detection of low abundance clonotypes and correction of amplification biases and sequencing errors. Furthermore, the dual-index amplicon labeling strategy minimizes index hopping during NGS allowing for comprehensive readouts.
While the DriverMap AIR-RNA assay quantifies T-cell and B-cell receptor transcripts (see Fig.2.) the DriverMap AIR-DNA assay amplifies receptor genes directly from genomic DNA. Combining data obtained from both the AIR-DNA and AIR-RNA assays enables assessment of both the transcriptional activation and number of cells with a particular clonotype. The ability to differentiate these two effects provides a quantitative basis to assess antigen-activated clonotypes.
Fig.1. Workflow of RNA/DNA assay.
To ensure consistent and reproducible T-cell Receptor (TCR) and B-cell Receptor (BCR) clonotype profiling Cellecta designed the DriverMap Adaptive Immune Receptor (AIR) RNA Spike-in Controls. A collection of synthetic mRNA constructs is offered that may be used as universal controls for commercial and home-brewed AIR sequencing assays based on multiplex RT-PCR or 5’-RACE (SMART) PCR techniques.
Fig.2. Number of clonotypes for 7 TCR/BCR chains identified in 50ng of normal PBMC or whole blood RNA (10x10^6 reads per sample, triplicates).
Do you have immune samples you would like to profile? Provide us with some information on your samples and your study goals, and we will get back to you to discuss your project.