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Overview

Understanding the intricacies of the immune system is crucial to elucidating roles of immune cells in health and disease. This requires profiling T- and B-cell receptors (TCRs and BCRs), key determinants of adaptive immunity. The next generation of BD Rhapsody™ TCR/BCR Multiomic Assays reveal immune repertoire richness with great sensitivity alongside simultaneous single-cell gene and protein expression analyses.

 
TCR-BCR Next product image
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Features

Validated on our new Enhanced Cell Capture Beads V3, BD Rhapsody™ TCR/BCR Next Multiomic Assays enable a highly sensitive analysis of full-length TCR/BCR repertoires with robust workflows for human and mouse. Key features include:

 

  • Superior full-length TCR and BCR pairing metrics
  • Multiomics-enabled for simultaneous immune profiling alongside protein and gene expression analyses in the same cell
  • Compatibility with preserved cells and BD® OMICS-Guard Sample Preservation Buffer
  • Intuitive data analysis pipeline that provides unbiased insights about the biology of your samples

 

Learn more from the BD Rhapsody™ TCR/BCR Next Multiomic Assay Brochure.

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Applications

A single workflow to identify and analyze T and B cell clonotypes with high sensitivity alongside single-cell whole transcriptome or targeted mRNA expression and protein analyses.

 

In this study, we simultaneously analyzed TCR, BCR, whole transcriptome gene expression as well as cell surface proteins using the BD® AbSeq Immune Discovery Panel in primary PBMCs. In total, 2,380 complete phenotypes were detected from 6,798 putative cells (out of 7,292 input cells).

Step 1:

Using both protein and gene expression, a UMAP plot was generated, and protein markers were used to identify cell populations.

 
UMAP Plot for Gene and Protein Markers
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Step 2:

An unsupervised analysis (phenograph) was performed on CD8+ cells where 11 clusters were identified.

 
11-Cluster CD8 Unsupervised Analysis
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Step 3:

Frequencies of unique clonotypes within CD8+ were reported.

 
Unique Clonotype Frequencies within CD8
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Step 4:

The top 5 clonotypes were overlaid on the UMAP plot and identified within phenograph clusters.

 
Clonotype Identification on UMAP Plot within Phenograph Clusters
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Step 5:

Using WTA and AbSeq information, differential gene and protein expression analysis of cells with the most frequent clonotype vs the rest of the population were performed.

 
Differential Gene and Protein Expression Cell Analysis with AbSeq and WTA
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Step 6:

 

Cell IDs and chain information were obtained from the pipeline output files and VDJ and C gene information were determined for the clonotype of interest.

 

Cell IndexTCR Alpha Gamma V gene dominantTCR Alpha Gamma J gene dominantTCR Alpha Gamma CDR3 translation dominantTCR Beta Delta V gene dominantTCR Beta Delta D gene dominantTCR Beta Delta J gene dominantTCR Beta Delta C gene dominantTCR Beta Delta CDR3 translation dominantTCR Paired ChainsCell type experimentalHigh quality cell
236563TRAV41*01TRAJ49*01AVRSGGDTGNQFYTRBV5‑6*01TRBD2*02TRBJ2‑1*01TRBC2ASSLREYNEQFTRUET_CD8_naiveTRUE
638393TRAV41*01TRAJ49*01AVRSGGDTGNQFYTRBV5‑6*01TRBD2*02TRBJ2‑1*01TRBC2ASSLREYNEQFTRUET_CD8_naiveTRUE
1789392TRAV41*01TRAJ49*01AVRSGGDTGNQFYTRBV5‑6*01TRBD2*02TRBJ2‑1*01TRBC2ASSLREYNEQFTRUET_CD8_naiveTRUE
2461787TRAV41*01TRAJ49*01AVRSGGDTGNQFYTRBV5‑6*01TRBD2*02TRBJ2‑1*01TRBC2ASSLREYNEQFTRUET_CD8_naiveTRUE
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Step 7:

 

Full-length sequence of the clonotype was obtained (nucleotide sequence also available) from the pipeline output.

 

 

Cell IndexVDJ Translation Trimmed
236563KNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRSGGDTGNQFYFGTGTSLTVIP
236563DAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLREYNEQFFGPGTRLTVL
638393KNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRSGGDTGNQFYFGTGTSLTVIP
638393DAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLREYNEQFFGPGTRLTVL
1789392KNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRSGGDTGNQFYFGTGTSLTVIP
1789392DAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLREYNEQFFGPGTRLTVL
2461787KNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGKKKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRSGGDTGNQFYFGTGTSLTVIP
2461787DAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLREYNEQFFGPGTRLTVL

 

 FR1  CDR1  FR2  CDR2  FR3  CDR3  FR4
TCR_alphaKNEVEQSPQNLTAQEGEFITINCSYSVGISALHWLQQHPGGGIVSLFMLSSGK KKHGRLIATINIQEKHSSLHITASHPRDSAVYICAVRSGGDTGNQFYFGTGTSLTVIP
TCR_betaDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLREYNEQFFGPGTRLTVL
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.