| Texto completo | |
| Autor(es): |
Sant'Ana, P. M.
[1]
;
Oliveira, J. E.
[1]
;
Lima, E. R.
[1]
;
Soares, C. R. J.
[1]
;
Peroni, C. N.
[1]
;
Bartolini, P.
[1]
;
Ribela, Maria Teresa C. P.
[1]
Número total de Autores: 7
|
| Afiliação do(s) autor(es): | [1] IPEN CNEN, Dept Biotechnol, Ave Prof Lineu Prestes 2242, Cidade Univ, BR-05508900 Sao Paulo, SP - Brazil
Número total de Afiliações: 1
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Applied Microbiology and Biotechnology; v. 102, n. 3, p. 1215-1228, FEB 2018. |
| Citações Web of Science: | 1 |
| Resumo | |
A strain of embryonic human kidney cells (HEK293) was transiently co-transfected with the expression vectors coding for the alpha- and beta-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 (t (1/2)) 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. (AU) | |
| Processo FAPESP: | 15/26058-0 - Estudo do perfil de N-glicosilação e dos sítios de ocupação em várias preparações de tireotropina humana (hTSH) hipofisária e recombinante |
| Beneficiário: | Carlos Roberto Jorge Soares |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |