Team:FAU Erlangen/Basic Part
Basic Parts
For the basic parts, components for the creation of bispecific antibodies for expression in E.coli and HEK 293T cells were provided.
Linker sequences
All the flexible linker are composed of small, non-polar (Glycine) and polar (Serine) amino acids. The Gly4Ser linker is one of the most widely used flexible linker that can sustain the bioactivity of fusion proteins. The distinct linker differ in the copy number of GGGGS, resulting in different bioactivities of the fusion proteins.
((Glycin)x4-Serine)x4 Linker)
(Glycin)x4-Serine Linker
(Glycin)x4-Serine Linker
Leader sequences
Igk is a leader sequence for the secretion of eukaryotic protein.
pelB is a leader sequence for targeting prokaryotic proteins into the periplasmic space.
single chain variable fragments
scFv VH targeting CD3, which was codon optimized for expression in CHO-S.
scFv VH targeting GPA33, which was codon optimized for expression in CHO-S.
scFv VL targeting CD3, which was codon optimized for expression in CHO-S.
scFv VL targeting GPA33, which was codon optimized for expression in CHO-S.
scFv VH targeting CD3, which was codon optimized for bacterial expression.
scFv VH targeting GPA33, which was codon optimized for bacterial expression.
scFv VL targeting CD3, which was codon optimized for bacterial expression.
scFv VL targeting GPA33, which was codon optimized for bacterial expression.
Constant regions for the creation of fabs
&&κ;;-light chain is the constant part of the light chain within an antibody.
The protein domain CH1 is the constant fragment of the heavy chain of an antibody.
Conjugation system
For conjugation, the SpyTag/SpyCatcher system was used in bacterial and eukaryotic expression, forming an isopeptide bond.
SpyTag for eukaryotic expression
SpyTag for prokaryotic expression
SpyCatcher for eukaryotic expression
SpyCatcher for prokaryotic expression