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Antigen-specific T cell staining with reversible MHC I Streptamers®

The MHC I Streptamers®, adducts of Strep-tagged MHC I molecules (MHC I-Streps) and Strep-Tactin®, are directed to antigen-specific CD8 T cell receptors.

Cell staining procedure

Cell staining procedure

1) Peptide (antigen)-loaded MHC I-Streps (Strep-tagged MHC I molecules) are multimerized with Strep-Tactin® attached to a fluorochrome to generate multimers with high binding avidity. These multimers are determined for staining of T cells which are specific for the respective peptide-loaded MHC molecule (Fig.1).

MHC I Streptamer fluorochrome
Fig. 1: MHC I Streptamer® Fluorochrome

2) After FACS analysis or FACS sorting of the target cells, the staining reagents can be completely removed with biotin (Fig.2). Cells are left phenotypically and functionally indistinguishable from untreated cells.

Fig.2: Antigen-specific cytotoxic CD8 T cells are selected by a MHC I Streptamer®. After staining and fluorescence assisted sorting, the Strep-Tactin® complex is removed from the target cells by the addition of biotin under physiological conditions at low temperatures (4°C). The monomeric low affinity MHC I-Strep molecules are then dissociated completely from the cell surface by two short washing steps. The isolated cells are phenotypically and functionally undistinguishable from non-treated cells and predestined for clinical use.

For further information about the antigen-specific T cell staining with MHC Streptamers® and the suitable reagents please have a look at our workflow.

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