Scott Hickey, the Director of Commercial Applications at Oxford Nanopore Technologies, spoke at the ONT Biopharma Day in Boston. The session’s title was “Antibody discovery through post-vaccination single-cell transcriptomics and haplotype-resolved germline sequencing.” Hickey spoke about a de novo full-length antibody identification through sequencing. B cell activation and proliferation were described along with the diversity of antibodies. Polymorphic loci and transcripts can be sequenced. For the case study, a blood sample was obtained from a person after the MMR vaccination. Genomic DNA and B cell RNA were sequenced. This entire experiment was done on a single PromethION flow cell. They obtained 16.4 M reads! A fully phased assembly of the IGH loci was obtained. Methylation signals of IGHV genes were analyzed. Single-cell transcriptomics was performed using 10x Genomics preps. The wf-single-cell EPI2ME workflow was then used for analysis. Hickey showed UMAPs of memory B cell subpopulations clustered by gene expression and isoforms. Full-length Ig consensus transcripts were then analyzed and compared between memory B cells and antibody-secreting cells. Hickey presented additional experiments to identify and synthesize antibodies against the virus! Thus, sequencing of B cell transcriptomes and genomes can help create personalized VDJC allele assignments!
