Fab Streptamers (lymphocytes) - Cell staining

Receptor-specific staining of lymphocytes with reversible Fab Streptamers

Fluorescent Fab Streptamers, adducts of Strep-tagged Fab fragments (Fab-Streps) and Strep-Tactin¢ç PE or APC, are directed against cell surface markers including those of  T cells (CD3, CD4, CD8), memory T cells (CD8, CD62L, CD45RA-) and regulatory T cells (CD4, CD25, CD45RA).

Cell staining procedure

1)    Fab-Streps (Strep-tagged Fab fragments) are multimerized with Strep-Tactin¢ç attached to a fluorochrome to generate multimers with high binding avidity. These multimers are directed to the respective cell surface marker(s) and are thus determined for staining of the target cell (Fig.1).
Fig.1: Fab Streptamer¢ç 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: Target cells are selected by a Fab Streptamer¢ç. After staining and fluorescence assisted sorting, the Strep-Tactin¢ç complex is removed from the target cells by the addition of biotin under physiological conditions at low temperatures (4¡ÆC). The monomeric low affinity Fab-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.

Fluorescent Fab Streptamers for reversible, receptor-specific staining of lymphocytes

For staining of cell isolates obtained after magnetic cell isolation with the Fab Streptamer¢ç isolation kits. Fab Streptamers for cell staining comprise a Strep-Tactin¢ç fluorochrome which has to be associated with a Fab-Strep in a short incubation step. The resulting fluorescent Streptamers are then used for marker-specific staining of the target cells. The Streptamers can be completely removed from the cells with biotin.
Fab_Streptamer_Fluorochrome.png
 
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