Antibody Variable Region Sequencing and Cloning

Curia has developed an efficient and reliable system that utilizes molecular cloning to sequence antibody variable regions (VH and VL) from hybridoma cell lines or clonal primary B cells across a variety of species.

As a quick overview, mRNA isolated from hybridoma samples is subjected to RT-PCR with unique species-specific primer sets to amplify the target regions. Antibody variable region sequencing (AVS) is Curia’s tried and true service with more than 5000 sequences identified. More than 95% of the samples have been successfully sequenced with at least one heavy chain and one light chain identified. Even with technically difficult samples, Curia’s approach has demonstrated a high success rate in numerous cases. Curia’s AVS service coupled with automated data tracking and management system provides customers with absolute confidence at every step.

Following the successful cloning of variable regions, the sequences can be fused to a variety of IgG backbones. Alternatively, variable region sequences can be reformatted as single-chain Fv (scFv) or bispecific antibodies in various configurations. Transiently produced antibodies can then be validated and characterized by downstream analytical services.