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DriverMap Adaptive Immune Receptor (AIR) TCR-BCR Profiling

DriverMap AIR is the only assay technology on the market that profiles the full or 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 RNA, DNA, or both. The simultaneous profiling enables the identification of antigen-activated clonotypes to provide even greater insight into the immune response.

How does the DriverMap AIR assay work?

  • Start from a variety of immune sample types such as whole blood, PBMCs, cancer biopsies, tissue samples, FFPE or dried blood microsamples.
  • Detect and accurate quantify AIR clonotypes with the multiplex PCR-based assay making use of Unique Molecular Identifiers (UMIs).
  • Obtain more sensitive and reproducible profiling data.

      • Why Do I Need Information on Both T-Cell and B-Cell Receptor Repertoires?

        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 provides the most sensitive detection of low-frequency rare TCR and BCR clonotypes and more comprehensive profiling when working with small samples and limited numbers of cells.

        The DriverMap AIR DNA-based assay provides a more quantitative measurement of the number of cells with each CDR3-specific clonotype. This data enables the measurement of clonal expansion in T and B cells.

        When information from both the AIR-DNA and AIR-RNA assays are combined, one can readily identify antigen-activated clonotypes. The heatmaps below (Fig. 1) show the activation of specific TRB and IGH clonotypes across multiple samples in metastatic tumor biopsies. For each sample (i.e., heatmap), data is normalized for RNA and DNA profiling; the green brackets show clonotypes present in RNA but not in DNA, indicating metastatic tumor-activated immune receptor clones

        Detection of Cancer-Activated CDR3 Clones
        Cellecta_DriverMap_Assay
        Fig. 1: Normalization of AIR-RNA to AIR-DNA profiling data identifies 5-10% of metastatic tumor activated immune receptor clones for TCR (β chain) and BCR (IGH chain) in multiple samples.

      • How is the DriverMap AIR Assay Different from other AIRR Assays?

        • 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 TCR/BCR isoforms.
        • Unique Molecular Identifiers (UMIs) facilitate accurate quantitation of the copy number of cDNA or DNA molecules in amplification steps, as well as detection of low abundance clonotypes and correction of amplification biases and sequencing errors.
        • Dual-index amplicon labeling strategy minimizes index hopping during NGS allowing for comprehensive readouts.
        • Full profiles of the antigen-recognition CDR3 region enable assessment of CDR3 length distribution, V(D)J segment usage, isotype composition for BCRs, somatic mutations, and similar characteristics with immune receptor profiling software such as MiXCR (MiLabs).
        DriverMap™ Adaptive Immune Receptor (AIR) TCR-BCR Profiling - Manual

      • Applications of BCR sequencing

        • Identify broadly neutralizing antibodies (BNAbs) and map Ig-seq datasets to known antibody structures for antibody and vaccine development.
        • Track B-cell migration and development patterns
        • Find markers of autoimmune diseases such as multiple sclerosis, rheumatoid arthritis and cancers (e.g. B-cell lymphoma)
        • Contrast naïve and antigenically challenged datasets to understand antibody maturation

        Applications of TCR sequencing

        • Track T-cell clonality and diversity for insights into mechanisms of action of immune checkpoint inhibitors for immunotherapies
        • Assess TCR overlap between repertoires to define spatial and temporal heterogeneity of the anti-tumoral immune response
        • Analyze TCR sequence and structure to annotate antigenic specificity for developing personalized cellular immunotherapies

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