Multiplex MHC multimer staining

For specialized purposes within detection of antigen-specific T cells ImmuMap offers analysis using the dCODE™ Dextramer platform.

Next-Generation Parallel Determination of multiple T cell specificities

This innovative platform takes advantage of unique DNA barcodes enabling us to search for up to more than 1000 different T cell specificities in a single cell sample using fluorescence-based cell sorting and next-generation sequencing (NGS). This method is based on the use of DNA barcoded MHC complexes and has been developed and pioneered in the lab of our co-founder Professor at the Danish Technical University, PhD Sine Reker Hadrup. The technology is essentially an expansion to the common MHC multimer staining, see our site about MHC multimer staining for an explanation of this method. We believe we are unique in offering the use of DNA barcoded MHC complexes as a service to our commercial customers.

What is DNA barcoded MHC complexes?

The dCODE™ Dextramer platform can be explained briefly as an initial standard staining of T cells with fluororescently-labeled peptide:MHC complexes followed by sorting of cells recognizing the peptide:MHC complex. This standard procedure is then followed by PCR amplification of the barcodes bound to the sorted cells and finally an NGS analysis. The dCODE™ Dextramers are, as common MHC multimers, built of a light chain, a heavy chain, a peptide and a fluorochrome-conjugated backbone. In addition, for each specificity the backbone has been labeled with a unique DNA barcode that can be sequenced and read by NGS.

All DNA barcoded MHC complexes are labelled with the fluorochrome PE and thus different T cell specificities cannot be elucidated from the flow staining alone. However, after sorting of the whole CD8+PE+ population we can identify the unique DNA barcode and consequently the specificity of the T cells in question by de-convoluting using next-generation sequencing and our own bioinformatics analysis platform.

Applicability of the dCODE™ Dextramer platform

The dCODE™ Dextramer platform can be explained briefly as an initial standard staining of T cells with fluororescently-labeled peptide:MHC complexes followed by sorting of cells recognizing the peptide:MHC complex. This standard procedure is then followed by PCR amplification of the barcodes bound to the sorted cells and finally an NGS analysis. The dCODE™ Dextramers are, as common MHC multimers, built of a light chain, a heavy chain, a peptide and a fluorochrome-conjugated backbone. In addition, for each specificity the backbone has been labeled with a unique DNA barcode that can be sequenced and read by NGS.

All DNA barcoded MHC complexes are labelled with the fluorochrome PE and thus different T cell specificities cannot be elucidated from the flow staining alone. However, after sorting of the whole CD8+PE+ population we can identify the unique DNA barcode and consequently the specificity of the T cells in question by de-convoluting using next-generation sequencing and our own bioinformatics analysis platform.

Learn more about
T-CELL RESPONSES

Learn how ImmuMap utilizes methods like Enzyme-Linked Immunospot (ELISpot), intracellular cytokine staining (ICS) and MHC multimer staining to characterize T-cell responses in innovative immunomodulating treatments.