THE SAME SIGNAL PEPTIDE IS NOT ALWAYS EFFICIENT

ATTALLAH CV, KRATJE RB, ETCHEVERRIGARAY M, OGGERO MR.

Buenos Aires

Congreso; Reunión anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2013

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB)

Most of the glycoprotein-based biotherapeutics are produced in mammalian cell lines because post-translational modifications should be identical or, at least, similar to those obtained in humans. Although CHO cells are the most prevalent for producing them, human embryonic kidney cells (HEK), and mouse myeloma (NS0) are being used as highproducing cell lines. On the other hand, signal peptides (SPs), capable of efficiently directing protein secretion in mammals, are key elements in recombinant protein production. In order to study the efficacy of three SPs to direct the secretion of a scFv-Fc fusion protein; CHO-K1, HEK293 and NS0 cell lines, producing the quimeric protein, were generated. In this way, the SPs derived from human albumin, human azurocidin and Cricetulus griseus Ig kappa chain V-III region MOPC 63 like were evaluated. The secretion efficiency of the fusion protein was analyzed by indirect specific ELISA from culture supernatant of each cell line. Although the best results were obtained with the human azurocidin derived SP for CHO-K1 and HEK293 cells, the scFv-Fc protein was not detected in the NS0 culture supernatant using the same SP. Nevertheless, the scFv-Fc protein was expressed by NS0 cells using the other two sequences. In conclusion, the human azurocidin signal peptide, being the best option for CHO-K1 and HEK293 cells, could not extrapolated to NS0 cells.