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Isolation of antigen-specific cytotoxic CD8 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 Isolation with Magnetic Nanobeads (for magnetic columns)

1) MHC I-Streps (Strep-tagged MHC I molecules) are multimerized with Strep-Tactin® attached to a magnetic bead to generate multimers with high binding avidity (Fig.1).

MHC 1 Streptamer magnetic nanobead
Fig.1: MHC I Streptamer® Magnetic Nanobead

2) After separation of the target cells on a magnet column, the isolation reagents can be completely removed from the cells with biotin (Fig.2). Cells are left phenotypically and functionally indistinguishable from untreated cells.

Fig.2: Streptamer® Magnetic Nanobeads are recommended when selection of cells is performed with an automated selection machine from Miltenyi (AutoMACS®, CliniMACS®). Cells are mixed with Streptamer® Magnetic Nanobeads and loaded onto a magnet column (Miltenyi). Cells bound to magnetic beads are retarded on the column through a high gradient magnetic field. After elution of the target cells multimerization is reversed by the addition of biotin and the low affinity Fab-Strep molecules dissociate completely from the cell surface.