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).

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.

Cell selection using MHC I Streptamer magnetic nanobeads

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.

**Magnetic MHC I Streptamers for antigen-specific isolation of (CD8+) T cells with reversible reagents**

Magnetic MHC I Streptamers comprise a MHC I-Strep which has to be associated with Strep-Tactin® Magnetic Nanobeads in a short incubation step. The resulting Streptamers are then used for antigen-specific isolation of the target cells on magnetic columns. After isolation, the Streptamers can be completely removed from the cells with biotin (stock solution included in the Streptamer® Solution Set Standard for washing and dissociation).

Modular Streptamer® technology: the MHC I-Streps can be associated either with a Strep-Tactin® fluorochrome for cell staining or with a Strep-Tactin® Magnetic Bead for cell isolation.

single MHC I Streptamer® products for magnetic cell isolation

Component	Use	Size
1) *MHC I-Streps*	choose among a great variety of different alleles and antigens	for staining of 2.5x10^8 cells or for isolation out of 5x10^8 cells human or 2.5x10^8 cells mouse*
2) Strep-Tactin® Magnetic Nanobeads	to be associated with MHC I-Streps	for isolation out of 5x10^8 cells human or 2.5x10^8 cells mouse*
3) Streptamer® Solution Set Standard	Buffer IS 10x for washing Biotin stock solution for dissociation	1 x 50 ml 1 x 1 ml
4) Streptamer® Solution Set XL	Buffer IS 10x for washing Biotin stock solution for dissociation	1 x 550 ml 1 x 5 ml
5) Nylon filter tubes	for removal of cell clumps	10 pieces

*For larger amounts please refer to the product list or inquire at info@streptamer.com

MHC I Streptamer® Magnetic Isolation Kits

Assemble your MHC I Streptamer® magnetic Isolation Kit in two steps:

choose your desired Isolation Kit; use filters "Category" and "Subcategory" on top of the product list
select your preferred MHC I-Strep

The kit includes magnetic nanobeads, MHC I-Streps and Solution Set Standard (Buffer IS for washing and Biotin for dissociation) for isolation out of 5x10^8 cells human or 2.5x10^8 cells mouse.

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