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DriverMap™ Adaptive Immune Receptor (AIR) Profiling Assay

DriverMap™ Adaptive Immune Receptor (AIR) Profiling Assay

Comprehensive, Sensitive, Reproducible AIR Profiling Without Specialized Equipment.
Available as a Kit starting from human DNA OR total RNA and as a Service

NEW!Now Actively Seeking Collaborators for Mouse TCR / BCR Profiling Assay
Profiling the T-cell receptor (TCR) and B-cell receptor (BCR) variable regions using RT-PCR and NGS can provide critical data useful for the discovery of novel, disease-associated immunity biomarkers and much more.

Did you know that from Cellecta’s DriverMap™ Adaptive Immune Receptor (AIR) Repertoire Profiling Assay you can

  • Obtain data on all 7 TCR/BCR isoforms from a single assay from total RNA (Fig. 1), or only TCR or BCR data from DNA or RNA
  • Amplify only functional CDR3 RNA molecules' TCR and BCR cells avoiding pseudogenes, ORFs, and the noise of non-functional sequences
  • Prepare Illumina-compatible next-generation sequencing (NGS) libraries simplyto get rapid results
  • Start with total RNA or DNA from any immune sample, including whole blood, FFPE, tissue biopsies, and more
  • Get antigen-activated clonotype info if you run DNA and RNA from the same samples
Fig. 1 Data from all 7 TCR/BCR chains from a single RNA sample.Chart shows the number of clonotypes for 7 TCR and BCR chains detected from 50 ng of PBMC and whole blood RNA samples in triplicate.
Adaptive Immune Receptor Repertoire Profiling
Fig. 2 Overview of the general structure of mRNA for different TCR (alpha, beta, gamma, delta) and BCR (heavy, kappa, lambda) chains and positions of the forward and reverse PCR primers to amplify the multiple CDR3 chains in a multiplex RT-PCR reaction 

The DriverMap™ AIR Assay Difference

More Clonotypes Detected by DriverMap AIR vs SMART RNA-based TCR Assay and Leading DNA-based Assay

  • Starting with the same amount of total RNA, the DriverMap™ AIR Assay identifies three-fold more clonotypes than the conventional SMART-based TCR assay (Fig. 3) and up to two times more clonotypes than the Adaptive immunoSEQ assay (Fig. 4).
DriverMap vs SMART
Fig. 3 Comparison of unique TCR clonotypes detected by DriverMap™ AIR vs SMART Technology. Both assays were run with 50 ng total RNA isolated from PBMC cells. DriverMap™ AIR detects ~3X more TCR clonotypes 
Fig. 4 Comparison of clonotypes detected by DriverMap™ AIR (mRNA) versus ImmunoSEQ (gDNA) assay from 21 tumor samples.DriverMap AIR Assay run on mRNA samples detects 1.5-2x more TCR and BCR clonotypes than gDNA samples run using the ImmunoSeq(R) assay.
Reproducibility is significantly higher in triplicate RNA samples with the DriverMap™ AIR assay for immune receptor profiling (iRP) when compared with the SMART-based assay (Fig. 5).  
Fig. 5 Comparison of the overlap and total number of all detected TRB clonotypes across triplicate parallel assays using DriverMap™ AIR and SMART immune profiling assays run with 50 ng of PBMC and whole blood RNA samples. NGS depth/replicate ~25M reads 
Robust Results with Small Samples Due to the low level of nonspecific background products, the DriverMap™ AIR Assay allows you to generate CDR3 profiling data from 10 ng to 1 ug of total RNA isolated directly from whole blood.

Furthermore, robust profiling data has been generated from just 30 ul of dried whole blood collected using a microsampling technology. In contrast, SMART technology is limited to use for immune profiling of PBMC RNA due to high background levels.

Uniform and Unbiased CDR3 Amplification DriverMap™ AIR assay amplifies shorter (300-400 bp) CDR3 fragments rather than full-length 700-800 bp V(D)J regions and so the amplification is more uniform and consistent(Fig. 6), though profiling with full-length sequences is also an option.
Fig. 6 Analysis of TCR transcript UMIs from DriverMap™ AIR and SMART assays. The DriverMap™ AIR assay produces a very consistent ratio of unique molecular indicators (UMIs) vs. reads for each transcript, whereas the SMART-based assay has a much less consistent ratio. With DriverMap™ AIR, amplification across all the transcripts was very consistent.

Combine Immune Repertoire and Gene Expression Profiling in the Same Workflow 

Both Cellecta’s targeted DriverMap™AIR andDriverMap™ Gene Expression Profiling Assays have the same workflow and ability to generate data from small quantities of total RNA. Both immune repertoire and gene expression profiles can be efficiently processed in parallel from the same total RNA samples, making it possible to obtain both RNA expression profile and immune repertoire profile data from the same samples.