Costruction And Expression Of Anti-HIV-1 ScFv Recombinant Diphtheria Toxin

At present, recombinate immunotoxin known as "biologic missiles" is extensively focused on the research of tumor, autoimmune disease, graft rejection and other immunotherapies. For AIDS, because immunotoxin can kill the potential infected cells that have been dormant infected by the HIV selectively and ruin the virus reservoir thoroughly, also being safe to the normal cells, it become highlighting on adjunctive therapy.In order to acquire ideal HIV drugs without side effects, we co-expressed ScAb and rIT in E.coli, and hoped that by the mediated effect of ScAb rITs can kill the HIV cells selectively. Therefore, the idea of targeting therapy is accomplished.To express ScFv containing chimeric immunotoxins , a ScFv expressing plasmids and two immunotoxins expressing plasmids by replacing the receptor binding domain or with transmembrane domain of DT genes with ScFv gene have been constructed. Plasmid pET-DT-hS120 and pET-DTA-hS120 were constructed, then transformed into E.coli strain BL21 (DE3) and induced by IPTG. Both of these two immunotoxins (DT-hS120, DTA-hS120) were expressed successfully. By one step of Ni-NTA affinity chromatography, and the recombinant immunotoxins was purified.