DriverMap AIR is the only assay technology on the market that profiles the full-length receptor region 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 RT-PCR and next-generation sequencing (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.
How does the DriverMap AIR assay work?
Since T- and B-cells work synergistically in the adaptive immune response, we have designed an assay that profiles both T-cell receptor (TCR) and B-cell receptor (BCR) repertoires in a single convenient reaction. Separate assays specific for T- or B-cell chains are also available.
The DriverMap AIR-RNA assay quantifies T-cell and B-cell receptor transcripts. Assaying expression of the immune receptors from either the CDR3 or the full-length receptor region enables highly sensitive detection of low-frequency, rare TCR and BCR clonotypes and more comprehensive profiling when working with small samples and limited numbers of cells. Fig. 1 shows that the DriverMap AIR-RNA assay is able to detect all seven TCR and BCR chains from a single sample of RNA.
The DriverMap AIR-DNA assay amplifies receptor genes directly from genomic DNA. The AIR-DNA assay provides a more quantitative measurement of the genetic copies for each CDR3-specific clonotype which correlates to the number of cells with that clonotype in that sample. This data enables the measurement of clonal expansion in T and B cells.
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. For example, it is evident from the data in Fig. 2, that particular TRB clonotypes, even when present in only a small portion of the cells in a population, can be highly up-regulated.
Fig 1: Number of clonotypes for 7 TCR/BCR chains identified in 50ng of normal PBMC or whole blood RNA (10x10^6 reads per sample, triplicates).
Fig. 2: The red intensity of each line in the paired heat maps shows the detection level of each TCR or BCR clonotype in each sample with the DriverMap AIR-RNA (R) and a competitor DNA (D) assay. By comparing the normalized level of clonotype expression as measured by the AIR-RNA assay with the frequency of each clonotype in the population as measured by the DNA-based assay, it is evident that 5-10% of the clones (indicated by green brackets) are strongly up-regulated even when they are only weakly detected at the DNA level, indicating they are not highly represented in the population.
How is the DriverMap AIR Assay Different from other Adaptive Immune Receptor Repertoire (AIRR) Assays?
Applications of BCR sequencing
Applications of TCR sequencing