PRJNA412649
Paired Heavy and Light Chain Immunoglobulin Reconstruction in Single-Cell RNA-Seq Data
We generated scRNA-Seq data for 20 human plasmablasts induced by seasonal flu vaccination.To reconstruct the paired heavy and light chain antibody genes, we developed several different in silico filtering strategies to enrich immunoglobulin transcripts, and then applied de novo assembly. This method successfully reconstructed productive IgH and IgL chains in 100% of cells analyzed. The accuracy of clonotype-assignment of individual cells was 94.5-98%, as validated by comparing BCR sequences reconstructed from NGS to those obtained by conventional single-B cell cloning methodology. This makes it possible to link the transcriptional programming of individual B cell clones at critical developmental stages with the eventual fate of the clonal lineage in vivo.