Human thyroid-stimulating hormone synthesis in human embryonic kidney cells and related N-glycoprofiling analysis for carbohydrate composition determination
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2018
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Applied Microbiology and Biotechnology
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A 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.
Como referenciar
SANTANA, 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. Applied Microbiology and Biotechnology, v. 102, n. 3, p. 1215-1228, 2018. DOI: 10.1007/s00253-017-8684-8. Disponível em: http://repositorio.ipen.br/handle/123456789/28451. Acesso em: 26 Apr 2024.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.