Efficient expression and purification of recombinant sphingomyelinase D from Loxosceles laeta in Lepidopteran larvae as a candidate for antiserum production

MC CALLUM, GREGORIO JUAN; ARGNANI PATRICIA; IGNACIO SMITH; ARREGUI MARIANA; ALEXANDRA TARGOVNIK; FINGERMANN MATÍAS; MARIA V MIRANDA

CABA

Congreso; 20Th World Congress of the International Society on Toxinology.; 2019

Expression was successful in both Spodoptera frugiperda and Rachiplusia nu larvae, obtaining higher yields for Spodoptera frugiperda. Good purification levels were achieved with cationic-exchange matrixes and higher still with IMAC. The resulting protein was not glycosylated and inoculation in rabbits demonstrated dermonecrotic lesions comparable to whole venom. Preliminary test to evaluate immunogenicity of the recombinant protein established less potency compared to whole spider venom, but further analysis are required. Thus, Sphyngomyelinase D represents a good example of the use of insect larvae as a small bioreactor for the production of recombinant proteins in a cost-effective and easily scalable process.