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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Human thyroid-stimulating hormone synthesis in human embryonic kidney cells and related N-glycoprofiling analysis for carbohydrate composition determination

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Author(s):
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]
Total Authors: 7
Affiliation:
[1] IPEN CNEN, Dept Biotechnol, Ave Prof Lineu Prestes 2242, Cidade Univ, BR-05508900 Sao Paulo, SP - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Applied Microbiology and Biotechnology; v. 102, n. 3, p. 1215-1228, FEB 2018.
Web of Science Citations: 1
Abstract

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)

FAPESP's process: 15/26058-0 - Glycoprofiling and N-glycosylation site occupancy of different human thyrotropin (hTSH) preparations of pituitary or recombinant origin
Grantee:Carlos Roberto Jorge Soares
Support type: Regular Research Grants