SJ-21-0039 – Primer Panel for T cell receptor amplification

  • a multiplex primer panel for highly sensitive amplification of TCRantibody sequences from single or few T cells
  • generates a singlepooled sequencing library from either 96 or 384-well plates
  • Well informationis stored in a DNA-barcode introduced via PCR and the resulting library iscompatible with any high throughput sequencing platforms.

Abstract

Researchers at St. Jude have created a multiplex primer panel for highly sensitive amplification of TCR antibody sequences from single or few T cells, which generates a single pooled sequencing library from either 96 or 384-well plates. Well information is stored in a DNA-barcode introduced via PCR and the resulting library is compatible with any high throughput sequencing platforms. It also can simultaneously amplify other genes of interest in the same well, and it can be easily automated with robotic systems. This system can be used to retrieve information about paired TCRs or/and protein expression at a single cell resolution in context of viral infections, anti-tumor immunity, and autoimmune diseases; and that information can be further used to develop TCR-based therapies and diagnostics. The most direct application of this system is decoding epitope-specific T cell responses. Also, it can retrieve cell phenotype information and could be used to get repertoires from larger cell samples. This invention enables a reduction in cost, time, and material; if you are interested in licensing this technology in one of these or any other fields, please contact us.

Advantages

Researchers at St. Jude have created a multiplex primer panel for highly sensitive amplification of TCR antibody sequences from single or few T cells, which generates a single pooled sequencing library from either 96 or 384-well plates. Well information is stored in a DNA-barcode introduced via PCR and the resulting library is compatible with any high throughput sequencing platforms. It also can simultaneously amplify other genes of interest in the same well, and it can be easily automated with robotic systems.

Figure 1. Possible experimental workflow overview (T cells stained with DNA-barcoded antibodies and/or MHC-multimers and are sorted into plates.

Potential Applications

This system can be used to retrieve information about paired TCRs or/and protein expression at a single cell resolution in context of viral infections, anti-tumor immunity, and autoimmune diseases; and that information can be further used to develop TCR-based therapies and diagnostics. The most direct application of this system is decoding epitope-specific T cell responses. Also, it can retrieve cell phenotype information and could be used to get repertoires from larger cell samples.

Contact Information

Name: Shawn A Hawkins

Email: shawn.hawkins@stjude.org

Phone: 901-595-5354