SANTANA, P.M.OLIVEIRA, J.E.LIMA, E.R.SOARES, C.R.J.PERONI, C.N.BARTOLINI, P.RIBELA, MARIA T.C.P.2018-02-062018-02-062018SANTANA, P.M.; OLIVEIRA, J.E.; LIMA, E.R.; SOARES, C.R.J.; PERONI, C.N.; BARTOLINI, P.; RIBELA, MARIA T.C.P. Human thyroid-stimulating hormone synthesis in human embryonic kidney cells and related N-glycoprofiling analysis for carbohydrate composition determination. <b>Applied Microbiology and Biotechnology</b>, v. 102, n. 3, p. 1215-1228, 2018. DOI: <a href="https://dx.doi.org/10.1007/s00253-017-8684-8">10.1007/s00253-017-8684-8</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/28451.0175-7598http://repositorio.ipen.br/handle/123456789/28451A strain of embryonic human kidney cells (HEK293) was transiently co-transfected with the expression vectors coding for the α- and β-subunits of human thyroid-stimulating hormone (hTSH), and, for the first time, a human cell-derived recombinant hTSH was synthesized and extensively characterized. The purification strategy involving two steps provided an overall yield of 55% and a purity level > 90%. The purified material (hTSH-HEK) was analyzed and compared to a CHO-derived recombinant preparation (hTSH-CHO) and to a pituitary-derived (hTSH-Pit) preparation. The three preparations showed an equivalent purity (> 95%) with a hTSH-HEK molecular mass 2.1% lower than that of hTSH-CHO and 2.7% higher than that of hTSH-Pit. Remarkable differences were found in the carbohydrate moiety, the lowest sialic acid content and highest fucose content being observed in hTSH-HEK. In vivo biological activity was confirmed for the three preparations, the hTSH-HEK bioactivity being 39 and 16% lower than those of hTSH-CHO and hTSH-Pit, respectively. The hTSH-HEK circulatory half-life (t1/2) was also shorter than those of hTSH-CHO (1.5-fold) and hTSH-Pit (1.2-fold). According to these findings, HEK-293-derived hTSH can be considered to be useful for clinical applications, in view as well of its human origin and particular carbohydrate composition.1215-1228openAccessthyroid hormoneskidneyshormonesstimulationsynthesisin vivomass spectroscopyanimal cellsglycoproteinsArtigo de periódico310210.1007/s00253-017-8684-8https://orcid.org/0000-0001-8194-5230https://orcid.org/0000-0002-7982-178975.0082.50