Next generation sequencing of antibody repertoires

Due to the size and duplication of the Ig loci, it has been impossible to quantify which V genes are used for recombination, and how, and why. We are using next generation sequencing assays (RNA-seq, ChIP-seq) to gain a high-resolution picture of underlying mechanisms, e.g. ncRNA transcription, at individual V genes and intergenic regions. 

We have also developed an NGS assay (VDJ-seq), that can interrogate recombined immunoglobulin gene repertoires at unprecedented depth and resolution and provides the first unbiased and quantitative assay of V(D)J recombination, which will be a powerful resource for other researchers. 

VDJ-SEQ: high throughput DNA-based unbiased quantification of antigen receptor repertoires

VDJ-SEQ: high throughput DNA-based unbiased quantification of antigen receptor repertoires

 

Bolland et al Cell Reports 2016; Chovanec et al,  Nature Protocols 2018

Two mutually exclusive local chromatin states drive Igh recombination

Two mutually exclusive local chromatin states drive Igh recombination

 

Bolland, Koohy et al, Cell Reports, 2016