Immune Receptor Profiling is a powerful tool for characterizing adaptive immune responses to cancer, autoimmune and infectious diseases, allergies, vaccinations,and therapeutic treatments. The unique sequences of the T-cell and B-cell receptors (TCRs and BCRs), and antibody variable regions (CDR3) that recognize foreign antigens define the individual differences in adaptive immune responses. Profiling the TCR and BCR variable regions using RT-PCR and NGS provides critical data for the discovery of novel, disease-associated immunity biomarkers.
Cellecta offers two types of DriverMap™ Adaptive Immune Receptor (AIR) repertoire profiling assays: AIR RNA and AIR DNA. Both assays are based on targeted multiplex PCR amplification and next-generation sequencing (NGS) analysis of variable T-cell receptor (TCR) and B-cell receptor (BCR) sequences using RNA and/or DNA purified from biological samples as the starting material. AIR RNA is our front-line recommended assay which provides the highest performance in immune receptor profiling. The AIR DNA assay can be used separately or in combination with AIR RNA, as discussed below.
- An easy-to-use protocol that allows comprehensive, unbiased immune repertoire profiling from small samples (e.g., sorted cells, FFPE, biopsy or blood microsamples) or larger bulk samples (e.g., whole blood or tissue biopsies).
- A novel, multiplex RT-PCR protocol that provides the highest sensitivity in the detection of functional or productive CDR3 clonotypes and also eliminates non-specific, off-target, and primer-dimer amplification products.
- Built-in Unique Molecular Identifiers (UMIs) to identify and correct amplification biases and sequencing errors for both the AIR RNA and AIR DNA assays. The UMIs accurately quantitate copy number of cDNA / DNA molecules used in amplification steps and detect low-abundance clonotypes above the background level.
- A dual-index amplicon labeling strategy that minimizes the potential NGS shortcoming of index-hopping when using the Illumina Paired-End kit for CDR3 sequencing.
- Open-source software such as MiXCR (preferred choice), IMGT/HighV-QUEST, IgBLAST, IMSEQ, LymAnalyzer software can be used for AIR assay NGS data analysis. Cellecta also provides custom data analysis services with MiXCR software. We can generate interactive reports that include comprehensive somatic mutation profiling across the antigen-recognition CDR3 region, accurate clonotype quantitation, CDR3 length distribution, V(D)J segment usage, and isotype composition for BCRs.
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