Part:BBa_K3117030:Design
scFv against CD3 with SpyTag codon optimized for CHOs
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 604
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.