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Fluorescent cell staining of antigen- or receptor-specific T cells

Streptamers®: fully reversible reagents for uncompromised staining intensitv and accuracy

Cell biology is reliant on the investigation of unmanipulated cells. IBA's Streptamer® technology is the only fully reversible method for fluorescent cell staining as well as magnetic cell isolation. It offers excellent staining intensities and can be removed from the cells by addition of biotin (vitamin H).

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Streptamer® - Key features

  • no stimulation of cells after cell separation
  • optimal preservation of cell effector function
  • improved viability of cells
  • high reproducibility

Streptamer® - Principle

Animation of T cell staining with fluorescent MHC I Streptamers®:

Fluorescent MHC I Streptamers® Fluorescent Fab Streptamers®
for fluorescent staining of antigen-specific CD8+ T cells, for e.g. FACS for fluorescent staining of T cells and subsets, after previous isolation with the magnetic Fab Streptamers®
The Streptamer® principle: full reversibility
Low affinity Strep-tagged Fab fragments or MHC molecules are multimerized with Strep-Tactin®, a streptavidin derivative, to generate multimers with high binding avidity.  Strep-Tactin® fluorochromes are used for cell staining / FACS isolation and Strep-Tactin® magnetic beads are used for magnetic cell isolation. Upon addition of low concentrations of d-biotin (Vitamin H), which competes with high affinity for the binding of Strep-tag to Strep-Tactin®, staining and isolation reagents dissociate rapidly from the cell surface.
This reversibility enables multiple cell stainings with different Strep-Tactin® fluorochromes as well as sequential positive cell selections with magnetic beads.
The Streptamer® reagents: a modular system

Fab-Streps or MHC-Streps are either combined with fluorescence- or bead-labeled Strep-Tactin® for cell staining or magnetic isolation, respectively. Biotin is offered as stock solution with the Streptamer® Solution Set which is used for removal of cell staining as well as cell isolation reagents. This modular reagent system enables the economic generation of multiple receptor- or antigen-specific staining and isolation reagents. All reagents can be purchased as single products or assembled in ready-to-use staining and isolation kits.

Streptamer® - Application data

Streptamers®: Full reversibility and excellent staining intensities

Streptamers® are fully reversible Zhang et al., 2016

Streptamers® can be completely removed from the surface of antigen-specific T cells, and staining intensities are the same like that of conventional tetramers.

Streptamer® is more accurate than Pentamer Ciáurriz et al., 2016

Streptamer® is more accurate than pentamer for the detection of CTLA*02:01-CMVpp65495-503 and can be used not only for the monitoring of early CTLA*02:01-CMVpp65495-503 reconstitution in immunosuppressed patients following allo-HSCT but also, in conjunction with its reversibility role, for the isolation of CTLA*02:01-CMVpp65495-503 for its future use in adoptive immunotherapy.

Streptamers® are superior to tetramers and pentamers

Streptamer® is more accurate than Pentamer Ciáurriz et al., 2016

Streptamer® is more accurate than pentamer for the detection of CTLA*02:01-CMVpp65495-503 and can be used not only for the monitoring of early CTLA*02:01-CMVpp65495-503 reconstitution in immunosuppressed patients following allo-HSCT but also, in conjunction with its reversibility role, for the isolation of CTLA*02:01-CMVpp65495-503 for its future use in adoptive immunotherapy.

Streptamers® showed the best T cell yield as well as the best level and stability of enrichment. Govers et al., 2012

Streptamers®, when compared with tetramers and pentamers, showed the best T cell yield as well as the best level and stability of enrichment of TCR enigneered T cells.

Streptamers® allow predicition of cell reactivity. Hombrink et al., 2013

MHC tetramer staining is not always predicitive for specific T cell reactivity whereas Streptamers® allow a correlation between functional and structural avidity.

Streptamers® preserve cell function Neudorfer et al., 2007

Streptamers® preserve cell function, whereas lytic efficacy and proliferation is impaired in the presence of conventional tetramers.