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Streptamers®: Full reversibility and preserved function of isolated cells

Reversible Streptamers® confer full protection against L. monocytogenes infection in mice (Knabel et al., 2002).

The reversible Streptamers® maintain the in vivo function of isolated cells. In contrast, conventional MHC multimers/tetramers signifcantly change the phenotype and function of stained T cells at physiological temperatures (Knabel et al., 2002).

Pretreatment of LLO91-99-specific T cells with conventional binding MHC tetramer reagents resulted in significantly reduced protection towards Listeria infection following adoptive transfer. In contrast, after complete removal of the Streptamers with biotin, the same number of cells conferred almost an identical degree of protection as compared with positive controls.

Without removal of the Streptamers® before adoptive cell transfer, the cells were reproducibly more effective than cells coated with conventional MHC tetramers. Please read the complete paper Knabel et al., 2002.

 

Reversible Streptamers® preserve proliferation capacity and functional status of CD8+ T cells (Wang et al., 2013)

Proliferation of CMVpp65-specific CD8+ T lymphocytes was preserved after selection with reversible Streptamers®, while it might have been altered with tetramers (Fig.3). Furthermore, the functional status as measured by activity (secretion of IFN-gamma) and cytotoxic effectiveness (secretion of granzyme B) remained very active when selected with Streptamers® as compared to tetramers (Fig.4).


Reversible Streptamers® preserve T cell receptor expression of CD8+ T cells (Zhang et al., 2016)

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.

Streptamers®: Uncompromised, authentic cells for research and diagnostics

The realization of a truly reversible labeling enables, for the first time, the collection of fully functional cells that are indistinguishable from untreated cells. This is in contrast to the unfavorable effects of conventional multimer techniques.

The Streptamer® technology is likely to advance basic cell research as an essential prerequisite for novel immunological medicines. Dirk H. Busch (et al.), the inventor of the Streptamer® technology, introduced the Streptamers in the June 2002 issue of Nature Medicine. He describes the Streptamer® approach as a method to isolate, for the first time, fully functional antigen-specific T cells. Experiments in mice demonstrate that the Streptamer® technology is capable to provide antigen-specific T cells, which can be efficiently applied in adoptive transfer protocols. Schmitt et al. recently reported that adoptive transfer and selective reconstitution of Streptamer®-selected cytomegalovirus-specific CD8+ T cells lead to virus clearance in patients after allogeneic peripheral blood stem cell transplantation.

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Fully reversible Streptamers® for basic research

Cell biology is reliant on the investigation of unmanipulated and uncompromised cells. Unlike state-of-the-art receptor specific cell isolation procedures, the Streptamer® approach is fully reversible, i.e. staining or isolation reagents are mildly and completely removed by biotin (vitamin H) at low temperatures which results in the following advantages:

  • no stimulation of cells after cell separation
  • optimal preservation of cell effector function
  • improved viability of cells
  • biotin is non-toxic to cells and harmless for (clinical) in vivo applications

Advantages of Streptamer® cell staining and isolation

  • functional characterization of cells since complete dissociation of reagents leaves cells phenotypically and functionally indistinguishable from untreated cells
  • mild and rapid dissociation of isolation reagents using biotin (vitamin H, which is non-toxic to cells and harmless in in vivo applications)
  • sequential positive cell isolations for e.g. isolation of pure cell subsets or to save valuable samples
  • simultaneous isolation of T cells with different HLA types and different antigen-specificity
  • modular reagent system for both cell staining and isolation
  • free combination of different Fab- and MHC I-Streps® with Strep-Tactin® fluorochromes or Strep-Tactin® magnetic beads for versatile and economic cell stainings or isolations
  • brilliant cell stainings due to high multimerization grade
  • FACS with fluorescently labeled Streptamers®
  • easy-to-handle cell purification with magnets
  • magnetic microbeads for cell isolation with a permanent magnet (e.g. IBA´s StrepMan Magnet)
  • magnetic nanobeads for cell isolation with magnetic columns
  • bead isolates can be ideally detected with Strep-Tactin® fluorochromes