Stability of budded virus of baculovirus in serum-free medium

EBERHARDT, I; GIORIA, V.; MICHELOUD, G.; CLAUS, J.D.

Potrero de los Funes-San Luis

Congreso; XLVII Reunión Anual de la Sociedad Argentina de Bioquímica y Biología Molecular; 2011

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Biotechnology applications of baculoviruses are constantly expanding. Any of these applications, from biopesticides to vectors in gene therapy, requires the proper production and preservation of high-titer stocks of budded virus (BVs) in serum-free culture medium. It has been described that frozen baculovirus storage in serum-free media is less efficient than in culture media added with serum, but the causes are unknown. The stability of BVs of the baculovirus AgMNPV was evaluated in a serum-free medium under different conditions of lipid supplementation, freezing and thawing, and exposure time to the production temperature (27°C), employing a 23full factorial design in duplicate. The time of exposure to 27°C, as wells as the freezing and thawing of samples, did not affect signifficantly the stability of BVs. However, it was strongly altered by the presence of lipids in the culture medium: the mean titer of samples preserved in medium supplemented with lipids was almost 4 times lower than the mean titer of samples stored in lipids-free medium. The deleterious effect of the lipids was magnified when BV samples were frozen and thawed: more than 90% of the mean viral infectivity was lost when samples supplemented with lipids were frozen and thawed. These results strongly suggest that the reduced stability of baculovirus BVs in serum-free media is associated to the presence of lipids.