Development of a production system for Z VLPs in insect cells

CRISTINA S BORIO; BILEN MARCOS; LOZANO MARIO ENRIQUE

San Luis

Congreso; Sociedad Argentina de investigación en Bioquímica y Biología Molecular; 2011

The assembly of proteins as virus‑like particles (VLPs) using protein expression technology in eukaryotic cells is a highly effective type of subunit vaccine, already tested and approved. The Arenavirus´s Z protein is involved in the assembly and budding of virions from the cell membrane. It also has been reported that the only expression of Z is sufficient for the release of Z-containing particles of Lassa Fever Virus. The generation of this VLPs from Z expression could have different biotechnological applications, such as drug and antigen delivery. Because of this is necessary the development of a safe, efficient and easy to scale up VLPs production system. Insect cells are a suitable candidate, since they express large amounts of correctly folded recombinant proteins and have been used for many previously approved vaccines as Cervarix. In order to determine if Junin virus Z protein maintains its budding capacity on insect cells, the recombinant Z-EGFP protein was expressed on Sf9 insect cells using the Bac-to-Bac system. We analyzed the culture supernatant of infected cells expressing Z-EGFP and detected particles preserved from degradation of proteinase K by a lipid membrane. These results might indicate the presence of Z-GFP in particles obtained from the culture supernatant.