Part Improvement:
We have worked to create an improved Glycine serine linker in part BBa_K1537017 in our new part BBa_K3244026
to adapt for our bispecific extracellular domain which should carry 2 scFv targeting 2 different antigens. This part has been modified through codon optimization -which has represented an issue for our fragment DNA synthesis due to the need for repetition of linker sequence between 4 different chains of 2 scFvs- and amino acid sequence length editing t(GGGGS)5 to act as an inter-chain linker between the 2 scFv for enhanced production and stability of the bispecific scFv. Significant difference between GSSS_ BBa_K1537017 engineered and GSSS_BBa_K3244026 T-cells(p<0.05), higher expression levels were observed inGSSS_BBa_K3244026 group.
group | Ct (EGR2ab) | Ct(ACTB) | Δ Ct (EGR2ab) | ΔΔ Ct(EGR2ab) | RQ(EGR2ab) |
T cells of BC associated Schistosomiasis patient (engineered with GSSS in BBa_K1537017 ) | 39.26 | 29.22 | 10.04 | 0.00 | 1.00 |
T-cells of BC associated Schistosomiasis patient BC associated Schistosomiasis (engineered with GSSS in BBa_K3244026) | 38.51 | 28.26 | 10.25 | 0.21 | 0.86 |
group | Ct (MEAab) | Ct(ACTB) | Δ Ct (MEAab) | ΔΔ Ct(MEAab) | RQ(MEAab) |
T cells of BC associated Schistosomiasis patient (engineered with GSSS in BBa_K1537017 ) | 40.26 | 29.22 | 11.04 | 0.15 | 0.90 |
T-cells of BC associated Schistosomiasis patient BC associated Schistosomiasis (engineered with GSSS in BBa_K3244026)" | 32.15 | 21.26 | 10.89 | 0.00 | 1.00 |