Part:BBa_K3117029:Design

scFv bispecific antibody against GPA33 and CD3 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 947
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Figure 1: Modeling of the 4x[GGGS] linker

The part consists of humanized anti-CD3 and anti-GPA33 single-chain variable fragments (scFv) (BBa_K3117020, BBa_K3117021, BBa_K3117022, BBa_K3117023), which are connected by a flexible [GGGGS]4 linker (BBa_K3117004). The scFvs are formed by the variable regions of the heavy and light chain of the respective antibodies that are connected by either a [GGGGS]4 or [GGGGS]3 linker (BBa_K3117004, BBa_K3117028). 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 domain 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.