195:. Therefore, a MHC multimer based on the interaction of Strep-tag with Strep-Tactin is easily disrupted in the presence of relatively low concentrations of d-biotin. Without the Strep-Tactin backbone, the single MHC-Strep-tag fusion proteins spontaneously detach from the TCR of the T cell, because of weak
214:
Knabel, M., Franz, T.J., Schiemann, M., Wulf, A., Villmow, B., Schmidt., B., Bernhard, H., Wagner, H. and Busch, D. (2002) Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer. Nature
Medicine 8 (6),
36:, that is conjugated to a marker which enables the isolation. The reversibility of this interaction and the low temperatures at which it is performed allow for the isolation and characterization of functional T-cells. Because T-cells remain
52:. They are capable of orchestrating, regulating and coordinating complex immune responses. A wide array of clinically relevant aspects are associated with the function or malfunction of T-cells:
179:
analysis. Incubation of MHC-Strep-tag fusion proteins with the Strep-Tactin backbone results in the formation of a MHC-multimer, which is capable for antigen-specific staining of T cells.
102:, which is the MHC-peptide complex, as a staining probe. The MHC interacts with the TCR, which in turn is expressed on the T cells. Because TCR-MHC interactions have only a very weak
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Schmidt TGM and Skerra A, 2007. The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. NATURE PROTOCOLS 2, 1528-1535.
122:. Nowadays, MHC molecules can be produced recombinantly together with the antigenic peptides which are known for a fast-growing number of diseases.
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MHC-epitope complexes cannot provide stable binding. This problem can be solved by using multimerized MHC-epitopes, which increases the binding
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and functionally indistinguishable from untreated cells, this method offers modern strategies in clinical and basic T-cell research.
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technology. In principle, the T-cells are separated by establishing a specific interaction between the T-cell of interest and a
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91:(MHC) procedures allow identification and purification of antigen-specific T cells independent of their functional status.
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has a much higher affinity to Strep-Tactin than Strep-tag, it can effectively compete for the
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molecule, called Strep-Tactin. For the
Streptamer technology, the Strep-Tactin molecules are
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conjugated to the MHC-multimers then can be used for identification of T cells by
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and form the "backbone", thus creating a platform for binding to strep-tagged
199:(monomeric MHC-epitope complexes cannot provide stable binding, see above).
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Strep Tamer is a technology which allows the reversible isolation and
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This technology combines a current T-cell isolation method with the
87:) have been developed for the identification of T cells, but only
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The
Streptamer staining principle combines the classic method of
151:-tag is a short peptide sequence that displays moderate binding
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versus host responses. Over the past years, various methods (
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171:. Additionally, the Strep-Tactin backbone has a
48:T cells play an important role in the adaptive
8:
94:In principle, MHC procedures are using the
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114:and therefore allows stable binding.
7:
44:Classic methods in T cell research
14:
89:major histocompatibility complex
245:from the original on 2004-09-18
81:intracellular cytokine staining
1:
279:Molecular biology techniques
159:-binding site of a mutated
305:
139:by MHC-multimers with the
126:The Streptamer technology
131:The Streptamer backbone
289:Laboratory techniques
187:Because the molecule
175:label to allow flow
106:towards each other,
183:Reversible staining
54:Autoimmune diseases
197:binding affinities
67:, development of
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147:technology. The
137:T cell isolation
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269:Protein methods
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96:T cell receptor
85:secretion assay
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38:phenotypically
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77:ELISpot Assay
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56:, control of
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50:immune system
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284:Biochemistry
247:. Retrieved
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193:binding site
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165:multimerized
161:streptavidin
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145:Strep-Tactin
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173:fluorescent
263:Categories
249:2023-08-13
203:References
24:-specific
177:cytometry
141:Strep-tag
108:monomeric
65:pathogens
62:bacterial
30:Strep-Tag
243:Archived
215:631-637.
189:d-biotin
169:proteins
155:for the
153:affinity
104:affinity
34:molecule
26:T-cells.
18:staining
112:avidity
22:antigen
235:"Home"
157:biotin
100:ligand
98:(TCR)
69:cancer
149:Strep
73:graft
58:viral
71:or
60:or
20:of
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