Targeted barcoding of variable antibody domains and individual transcriptomes of the human B-cell repertoire using Link-Seq.

ABSTRACT: Here, we present Link-Seq, a highly efficient droplet microfluidic method for combined sequencing of antibody-encoding genes and the transcriptome of individual B cells at large scale. The method is based on 3' barcoding of the transcriptome and subsequent single-molecule PCR in droplets, which freely shift the barcode along specific gene regions, such as the antibody heavy- and light-chain genes. Using the immune repertoire of COVID-19 patients and healthy donors as a model system, we obtain up to 91.7% correctly paired immunoglobulin heavy and light chains. Furthermore, we map the V(D)J usage and obtain sensitivities comparable with the current gold-standard 10× Genomics commercial systems while offering full flexibility in experimental setup and significant cost savings. A further uniqu

SUBMITTER: Hu H 

PROVIDER: S-EPMC11759286 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature