Part:BBa_K3398005:Design
scFv_Fc fusion protein
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1074
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 741
Design Notes
We employed codon optimization to maximize the expression of a functional protein in Escherichia coli. And most importantly, As the redox potential of general E.coli such as BL21 is too high to form the disulfide bond in scFv, which may not express the functional fusion proteins. So we chose E.coli Shuffle with a reduced cytoplasmic environment as the chassis organism to express scFv_Fc fusion protein.
Source
The amino acid sequence of anti-HER2 scFv and the CH2 & CH3 region of immunoglobulin heavy constant gamma 1, which is the constant region of immunoglobulin heavy chains (UniProtKB-P01857)(https://www.uniprot.org/uniprot/P01857), was used respectively.In addition, the hinge region of immunoglobulin heavy constant gamma 1 was used as the linker between scFv and Fc.