Forward icon

iPair: Single cell mPCR/NGS paring TCR alpha and beta chains

iRepertoire is proud to announce a new service called iPair-TCR. With iPair-TCR, the near full length sequences of human TCR alpha and beta chain genes are amplified from a single cell, barcoded, and sequenced by NGS. No need for computer algorithms, no guess work required. It is the first commercially available service open to all researchers, academic or industry alike. As usual, we do not claim reach through IP rights— you are the inventor of the content, and you own the IP for future diagnostics or drugs. We are here to help you achieve your research goals. Just ship in the samples, and we do the rest.

Since 2007, iRepertoire has been developing technologies and products that use NGS to study T and B cell repertoires. We were the first to file and obtain critical patent in this field (Patent 9012148). We have developed products, services and software for your research and development needs. However, when using existing methods, information related to the cognate pairing of alpha and beta chains is lost once RNA /gDNA extraction is performed on the entire sample (or bulk sample).

Traditional single cell RNAseq may recover the pairwise receptor information; however, since one T cell may differ from another only by 20-30bp in the CDR3 region, sequencing the entire transcriptome from a single cell just to pair the two chains is challenging in both technical and economical terms.

What distinguishes iRepertoire from other competitors is our powerful multiplex PCR technology called arm-PCR (amplicon rescued multiplex PCR). Using arm-PCR to generate targeted sequencing libraries, iPair-TCR can achieve greater than 90% success rate in single cell all-in-one amplification. Pairing can be confirmed in approximately 75% of these cells. Most of the “failed” cells may due to biological reasons such as gamma-delta T cells, or dual-alpha or beta chains expressed in one cell.

To ensure broader access to this powerful iPair-TCR technology, we have also developed comprehensive reagent systems and standard protocols for sample collection, storage, international shipping, sorting, and single cell arm-PCR amplification. Single cell plating is performed with a Sony SH800 FACs. Need additional sorting services? No problem, we are happy to provide options when customizing your research project. A user friendly desktop software is also available, free of charge, for iPair data browsing and analysis. In-house iPair-TCR amplification kits will be available at a later date, so stay tuned.

If you are interested in the service we would be happy to set-up an appointment to discuss your custom project needs.  Please email Customer Service at info@irepertoire.com for more information.  Otherwise, to access a demo of the iPair Analyzer software, please follow this link.

iPair-TCR+30: Single cell mPCR/NGS paring alpha & beta chains and profile 30 important genes

In addition to pairing alpha and beta chains from a single cell, iPair-TCR+30 enables us to study the expression of 30 important genes that define the functions of a T cell (genes listed in the table above).

This service is open for early access customers and will be fully available to everyone in March of 2017. For the iPair-TCR+ line of service, we are adding more genes to the list for profiling. We welcome project discussions and your suggestions for the addition of genes of interest to you.

Whats next? B cell? mouse? Others?

Yes, iPair-BCR for human B cell is the next service we plan to launch. Similar to iPair-TCR, B cell heavy and light chain genes will be amplified from a single cell for NGS. This will become a powerful tool for monoclonal antibody development. Instead of hybridoma technology, human mAbs can be developed directly by NGS by comparing immune repertoires before and after antigen exposure (PCT/US2012/058128).

Although iPair-TCR and iPair-BCR are only available initially for human samples, we are developing similar services for mouse as well. We are also developing exciting high throughput pairing methods using microfluidic devices for higher cell count samples.

Looking forward to hear from you in 2017. Happy New Year!