Difference between revisions of "Part:BBa K3117030:Design"

 
 
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===Design Notes===
 
===Design Notes===
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By connecting the variable regions of the heavy and the light chain of an anti-CD3 antibody with a short GGGGS linker (<partinfo>BBa_K3117004</partinfo>), the scFv retains it's antigen-binding ability and is much smaller than a conventional antibody. Thus, it is well suited as part of a fusion protein with another effector. The SpyTag attached to the scFv belongs to the SpyTag/SpyCatcher system and is one part of the FbaB protein of Streptococcus pyogenes. Once it comes into contact with its corresponding other part, the SpyCatcher (<partinfo>BBa_K3117016</partinfo>), they bind covalently. This allows our part to be used in a modular manner in combination with other molecules carrying the SpyCatcher. The sequence contains a C-terminal His-tag (<partinfo>BBa_K3117005</partinfo>) for easy purification and detection (Schmitt et al., 1993). Secretion of the protein is ensured by an Igk leader (<partinfo>BBa_K3117006</partinfo>). When the protein passes the membrane, this leader segment is cleaved off.
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===Source===
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===References===
 
===References===
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1. Schmitt, J., Hess, H., & Stunnenberg, H. G. (1993). Affinity purification of histidine-tagged proteins. Molecular biology reports, 18(3), 223-230.

Latest revision as of 22:18, 21 October 2019


scFv against CD3 with SpyTag codon optimized for CHOs


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 604
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

By connecting the variable regions of the heavy and the light chain of an anti-CD3 antibody with a short GGGGS linker (BBa_K3117004), the scFv retains it's antigen-binding ability and is much smaller than a conventional antibody. Thus, it is well suited as part of a fusion protein with another effector. The SpyTag attached to the scFv belongs to the SpyTag/SpyCatcher system and is one part of the FbaB protein of Streptococcus pyogenes. Once it comes into contact with its corresponding other part, the SpyCatcher (BBa_K3117016), they bind covalently. This allows our part to be used in a modular manner in combination with other molecules carrying the SpyCatcher. The sequence contains a C-terminal His-tag (BBa_K3117005) for easy purification and detection (Schmitt et al., 1993). Secretion of the protein is ensured by an Igk leader (BBa_K3117006). When the protein passes the membrane, this leader segment is cleaved off.

References

1. Schmitt, J., Hess, H., & Stunnenberg, H. G. (1993). Affinity purification of histidine-tagged proteins. Molecular biology reports, 18(3), 223-230.