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BCR Sequencing of 1 Million Healthy and Diseased Samples in a Single Experiment

Key Takeaways

  • 1 million human B cells in a single experiment
  • 900,000+ unique paired clonotypes detected from Type-1 Diabetes, Multiple Sclerosis, Rheumatoid Arthritis, Crohn’s and Celiac, Healthy donor samples
  • Sensitive CDR3, clonotype, and full-length detection at scale

Evercode BCR uncovered over 900,000 unique paired clonotypes across 24 samples in a single experiment.

Negatively selected B cells from 12 healthy donors were purchased. Pan B cells were isolated from 12 auto-immune diseased human PBMCs and fixed using Evercode Cell Fixation Kit v3 to stabilize the cell structure and protect the RNA transcriptome from degradation. Fixed samples were stored at -80C until all samples were ready for further processing in a single experiment with the Evercode BCR Mega kit. Whole transcriptome and BCR specific libraries were sequenced on Illumina Novaseq X, respectively. The data was analyzed using Parse Biosciences’ Analysis Pipeline v1.3.0.

Clustering of cells using Seurat 5.0 resulted in the majority of cells corresponding to the major B cell subtypes as demonstrated in the UMAP below (Figure 1).

Figure 1: UMAP clustering of all cells from 24 samples in a single experiment. All major B cell subtypes and other immune cells were detected.

The assay demonstrated high sensitivity, detecting paired heavy and light chains in up to 89% of cells (Figure 2).

Figure 2: BCR Chain Detection Across Six Sample Groups. High rates of chain assignments across the sample groups. Paired CDR3 detection varied between 54-89%.

Sample Table

SampleSexAgeEthnicityBlood TypePrepDisease state
H-1Male63HispanicO PosNeg select B cellsHealthy
H-2Male24African American/Native AmericanO PosNeg select B cellsHealthy
H-3Male73CaucasianO PosNeg select B cellsHealthy
H-4Female26Hispanic/LatinoA PosNeg select B cellsHealthy
H-5Female59HispanicA PosNeg select B cellsHealthy
H-6Male49Hispanic/MexicanO PosNeg select B cellsHealthy
H-7Male58African AmericanB PosNeg select B cellsHealthy
H-8Male64African AmericanA PosNeg select B cellsHealthy
H-9Male43African americanO PosNeg select B cellsHealthy
H-10Male21African AmericanO PosNeg select B cellsHealthy
H-11Male50CaucasianO PosNeg select B cellsHealthy
H-12Male33African americanB PosNeg select B cellsHealthy
MS-1Male33Hispanic/LatinoO PosPBMCMultiple Sclerosis
MS-2Male49CaucasianA PosPBMCMultiple Sclerosis
RA-1Female41CaucasianO PosPBMCRheumatoid Arthritis
RA-2Female50CaucasianO PosPBMCRheumatoid Arthritis
CD-1Female58CaucasianA NegPBMCCrohn's
CD-2Female22CaucasianO PosPBMCCrohn's
T1D-1Female33CaucasianO PosPBMCType-1 Diabetes
T1D-2Female36CaucasianA PosPBMCType-1 Diabetes
T1D-3Female54CaucasianA PosPBMCType-1 Diabetes
T1D-4Male49CaucasianO PosPBMCType-1 Diabetes
T1D-5Female72CaucasianO PosPBMCType-1 Diabetes
CE-1Male32CaucasianA NegPBMCCeliac


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