Adaptive immune repertoire sequencing ('Rep-Seq') is increasingly being used to evaluate the immune system, interventions targeting the immune system (such as vaccination) [1], and immune-derived therapeutics (e.g., immunotherapy and antibodies) [2]. For antibody discovery, repertoire sequencing has become an indispensable technology capable of generating vast pools of data from which to identify and characterize the best candidates using tools such as IGX Platform [3]. In this webinar, we describe our journey of setting up a validated Rep-Seq assay for BCR/Antibodies. Given the biological characteristics of the adaptive immune repertoire, important conditions for developing a high-quality Rep-Seq assay include: well-curated germline databases, high-quality sample prep and sequencing, and rock-solid processing software. To validate sequence processing and analysis software, we make use of a simulation framework that takes in an AIRR-compatible repertoire and outputs highly realistic HTS validation data. Furthermore, ENPICOM teamed up with Viroclinics/DDL (a Cerba company) to set up a reliable and robust high-quality Rep-Seq method. By designing and including spike-ins in pilot sequencing runs, we are able to better assess the characteristics of our Rep-Seq assay. By integrating both wet- and dry-lab development and testing, we provide a robust Rep-Seq quantification solution that makes use of the latest developments in genomics, bioinformatics, and drug development.
References:
[1] Arnaout et al, The Future of Blood Testing Is the Immunome; Front Imm, 2021, doi: https://doi.org/10.3389/fimmu.2021.626793
[2] Oude Lohuis et al, High-throughput Sequencing Technologies are Revolutionising Antibody Discovery; IBI, 2021. https://tinyurl.com/yrx637kp
[3] Technologies, challenges, and applications of immune repertoire sequencing; ENPICOM 2020, https://enpicom.com/download-white-paper-applications/