Quantitative gDNA TCR/BCR Sequencing with RepSeq IQ™
Obtain quantitative, precise T-cell and B-cell gDNA clonotype and cell frequency data across clinical research studies
The human adaptive immune system produces a vast array of VDJ receptor sequences, enabling tailored responses to diverse antigens. By accurately quantifying and analyzing this diversity, researchers can gain deeper insights into immune cell function, disease mechanisms, cell manufacturing production, and cell therapy response. However, comparing and analyzing Adaptive Immune Receptor Repertoire (AIRR) data from different clinical samples is challenging due to the lack of standardized controls.
Our new assay, RepSeq IQ, overcomes this challenge by placing immune repertoires on a standardized scale across samples, allowing for reliable quantification of clone frequencies and thorough characterization of unique T and B cell clones within each sample, paving the way for more accurate and insightful immune repertoire studies from gDNA.
Confidently analyze and interpret AIRR-seq clinical and cell engineering data with RepSeq IQ
Standardize clinical data across studies
Determine the total quantity of T and B cells in a sample population and the frequency of each clone to standardize patient data across clinical studies.
Quantitate and track pathogenic clonotypes
Track malignant or autoreactive T and B cell clones at the cellular level during immuno-oncology or autoimmune clinical trials.
Validate iPSC transduction
Characterize non-expressing constructs during induced pluripotent stem cell (iPSC) development.
Validated AIRR data for clinical data standardization and comparison
RepSeq IQ uses our direct-multiplex PCR chemistry to precisely amplify human TCR beta or BCR chains from gDNA templates, internal quantitative controls, and housekeeping genes. This method translates NGS reads into absolute copy numbers and quantifies the total number of cells sampled in each library, enabling standardization between samples, conditions, and study cohorts.
RepSeq IQ features:
- Quantitative: Determine the true percentage of T and B cells in an unsorted sample.
- Sensitive: Quantify individual clones at the cellular level.
- Inclusive: BCR heavy and light chain together in a single assay.
- Secure: Complete and exclusive access to raw and processed data.
- Adaptable: Compatible with PBMC, BMNC, PAXgene, and EDTA Whole Blood.
Easily quantify and track T and B cell clones across samples
A testing framework was established to test the ability of the assay to count total nucleated cells and calculate the percentage of any given clone.
In this framework, B-cells were depleted from a PBMC sample. B-cell depleted gDNA was then mixed with purified B-cell gDNA at specific ratios: 0%, 25%, 50%, 75%, and 100%. The 3-chain multiplex PCR assay with HKGs and IQC were used to amplify all contrived specimens. Data were analyzed by mapping the VDJ (both functional and non-functional) and identifying the reads for the IQC and HKG. This process was also performed for TCR quantification.
Treemaps depict the immune repertoire with each V-J-uCDR3 represented by a rounded rectangle. Dominant 300 clones from tracking mix tracked through 10-fold dilutions. Top panel shows the entire sample diversity including both the background and trackable clones. Bottom panel shows the trackable clones in color with the background clones in gray.