IAL   21557
INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Lack of stability of the infectivity of budded virus of Anticarsia gemmatalis multiple nucleopolyhedrovirus in serum-free medium supplemented with lipid microemulsions
Autor/es:
EBERHARDT I.; GIORIA V.; MICHELOUD G.; CLAUS J. D.
Lugar:
Buenos Aires
Reunión:
Congreso; 2012 International Congress on Invertebrate Pathology and Microbial Control and 45th Annual Meeting of the Society for Invertebrate Pathology ; 2012
Resumen:
Biotechnological applications of baculoviruses
are constantly expanding, from bioinsecticides to protein expression and gene
therapy vectors. The development of feasible processes for the production of
baculovirus and baculovirus-based products in insect cell cultures requires
that high-titer stocks of budded virus (BVs) being prepared and stored in
serum-free medium. It is known that the preservation of baculovirus BVs by
freezing at ultra-low temperatures in serum-free medium is less efficient than
in culture media added with serum, but the causes were not elucidated. The aim
of this work was to study the stability of the infectivity of BVs of the Anticarsia
gemmatalis multiple nucleopolyhedrovirus (AgMNPV) in the UNL-10 serum-free
medium under different conditions of
supplementation with lipid microemulsions, freezing at -80°C, and time
of exposure to 27°C, employing a 23 full factorial design in
duplicate. The exposure to 27°C, as well as the freezing and thawing of an
AgMNPV stock, did not affect the stability of the viral titer determined
through an end-point dilution method. On the other hand, the exposure to lipid
microemulsion provoked a significant negative effect on the stock titer. This
deleterious effect was magnified when BV samples in medium supplemented with
lipid microemulsion were frozen and thawed, losing more than 90% of the viral
infectivity. These results strongly suggest that the reduced stability of
AgMNPV BVs in serum-free media is associated to the presence of lipid
microemulsions, and indicate the necessity to establish alternative protocols
to store BVs stocks produced in serum-free medium supplemented with lipid
microemulsions.