DATE:  September 10, 2019

TIME:   8:00am PT

Next generation sequencing of the immune repertoire allows detailed, sequence-specific insight into the immune system’s adaptive response to environmental challenges. This information can be used for a variety of applications ranging from immune diversity assessments to developing immune therapies. Here, we present novel technologies for interrogating the immune repertoire from single cell to bulk RNA. We discuss the application of a single cell technology, iPair, to the physical cognate pairing of immune receptor chains from T and B cells. This approach allows for rapid and precise discovery of specific adaptive immune responses to an antigen. Utilizing the insights gained from single cell, we also present the application of a novel PCR technique, dimer-avoided-multiplex PCR (dam-PCR), to the all-in-one repertoire amplification of BCRs and TCRs in a single, quantitative multiplex reaction. Immune repertoire analysis when applied to bulk RNA typically focuses on one of the receptor chains, such as TCR-beta, while information of all receptor chains from one sample are desired for better understanding the immune response. Specifically, we were able to apply this method to the amplification of both FFPE and PBMC RNA using a multiplex primer system covering all TCR and BCR loci including TCR-beta, TCR-alpha, TCR-delta, TCR-gamma, BCR-IgH, and BCR-IgKL.  The ability to inclusively profile immune-specific RNA from FFPE samples provides the opportunity to sample historical repertoire data even in situations where sample RNA may be both limited in quantity and degraded in quality. 

Learning Objectives:

• Learn how SPRIbead selection can increase the sensitivity of targeted sequencing applications.

• Understand paired VDJ receptor information directly obtained from single cells provides actionable information that can be screened for activity.

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