HEK293 cell line: a promising host for the production of a hyperglycosylated hIFN‑α2b

AGUSTINA GUGLIOTTA; NATALIA CEAGLIO; BRENDA RAUD; GUILLERMINA FORNO; LAURA MAURO; MARINA ETCHEVERRIGARAY; RICARDO KRATJE; MARCOS OGGERO EBERHARDT

Villa General Belgrano

Simposio; GlycoAR 2016; 2016

CONICET-UNC-INSTITUTO LELOIR-IBYME-SOC LATIN GLICOBIOLOGÍA

Glycosylation is one of the most important post-translational modifications that proteins can undergo because it defines their biological and physicochemical characteristics. Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in different hosts.Chinese Hamster Ovary (CHO) cells are the most commonly used mammalian hosts for the production of commercially approved biopharmaceuticals. Human Embryonic Kidney (HEK) cells have gained importance due to their ability to produce glycoproteins that are very similar to the human naturally secreted proteins in terms of post‑translational modifications, specifically glycosylation. Another advantage is that this system avoids the expression of antigenic glycoforms like N‑glycolylneuraminic acid (Neu5Gc) present in CHO-derived products. For these reasons, there is an increasing interest in using human cell lines for the production of recombinant therapeutic proteins.In our laboratory, a hyperglycosylated IFN-α2b analog has been generated. It has been called IFN4N since it has four potential N-glycosylation sites which were introduced by point mutations in its protein sequence. As a result of the increment of the glycosylation degree, IFN4N showed increased molecular mass and charge in comparison with the unmodified molecule, resulting in an improvement in its pharmacokinetic properties. Despite IFN4N has lower in vitro activity, it showed a better performance in in vivo animal models of prostate cancer in comparison with the non-glycosylated IFN.The benefits demonstrated by IFN4N prompted us to develop CHO-K1 and HEK293 producer cell lines in order to investigate the differences in the carbohydrate composition of both products as well as its influence on their biological activity and pharmacokinetic properties.