Performance of Evercode™ BCR v4 in Human B Cells

Key Takeaways:

• Paired BCR and whole transcriptome profiling of human B cells
• Whole transcriptome analysis shows different B cell states
• High BCR CDR3 heavy and light chain pairing and full-length chain recovery in single cells

Experimental Design:

Single cell B cell receptor profiling paired with whole transcriptome analysis enables high-resolution characterization of B cell populations, including clonal identity and transcriptional state.

To evaluate the performance of Evercode BCR v4, cryopreserved human CD19+/CD27+ memory B cells and CD19+/CD27- naive B cells were obtained from a third-party provider. Aliquots from each sample were fixed and processed using the Evercode BCR v4, Human VDJ + WT workflow, generating whole transcriptome and BCR libraries were generated from the same cells. Libraries were sequenced on an Illumina NovaSeq instrument and analyzed using the Parse Biosciences analysis pipeline.

Whole transcriptome analysis demonstrated robust gene detection, and unsupervised clustering identified different B cell states, including naive, transitional, memory, proliferating, and rare plasmablasts.

Figure 1. Human B Cells Profiled with Evercode BCR v4. UMAP visualization highlights the detection of multiple B cell states.

Evercode BCR v4 demonstrated high sensitivity, detecting BCR light chains in 95% of cells and heavy chains in 96% of cells, with paired heavy and light chain BCR CDR3 in 91% of B cells assayed. The majority of heavy and light chains recovered were full length, with 79% and 76%, respectively.

Figure 2. Paired and Full-Length Heavy/Light Chain Recovery with Evercode BCR v4. High recovery of heavy, light, and paired CDR3 regions (A) and full-length heavy and light chains (B).

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