Evaluation of B cell receptor profiling platforms and analysis tools using next-generation sequencing data (BCR-SEQC). Homo sapiens

The goals of this study are to establish protocols for development of reference samples and data sets for the evaluation of reconstructing B cell receptor repertoires with NGS and to develop materials that can be utilized for performance bench-marking studies. We performed a pilot study to prove the concept of using contrived samples with two B-cell lines and PBMC cells pooled from three individuals and demonstrated the performance characteristics of seven BCR-seq assays with short and long reads technologies. Subsequently, we performed whole-genome sequencing, RNAseq, and BCR-seq on 50 B-cell lines to determine the clonotype, full-length transcript and expression abundance of BCR genes in each cell line. Cell lines that were determined unambiguously as monoclonal of BCR expression of both heavy and light chains can be used to generate reference materials for performance bench-marking studies by the research community.