Team:NCTU Formosa/Parts
To display the antibody on the E.coli outer membrane, we used Lipoprotein-Outer membrane protein A (Lpp-OmpA). According to the paper reference [1], We chose the first 9 amino acids of Lpp to be the signal peptide, and the 46-159 amino acids of OmpA to be the anchor, the C-terminally of Lpp-OmpA then fused the single chain variable fragment (scFv). We added a NcoI restriction side between OmpA and scFv so that we can change any scFv DNA sequence just by NcoI restriction enzyme.
By ligating the constitutive promoter (BBa_J23101), strong ribosome binding site (BBa_B0034) and Lpp-OmpA-scFv, we were able to display scFv on the E.coli outer membrane continuously. At the back of this part, we have added fluorescent proteins as the reporters.
In our current work, we chose three targeted drugs, Avastin (Bevacizumab, anti-VEGF)[2], Erbitux (Cetuximab, anti-EGFR)[3] and Herceptin (Trastuzumab, anti-HER2)[4] from Drugbank, selecting their single chain variable fra