Staining of antigen-specific MHC I T cells 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
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).
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.