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

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===Design Notes===
 
===Design Notes===
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The sequence contains a C-terminal His-tag for easy purification and detection. Secretion of the protein is ensured by an Igk leader. When the protein passes the membrane, the leader segment is cleaved off. 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 its 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.
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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.