Expression of glycosylated human prolactin in HEK293 cells and related N‑glycan composition analysis
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Prolactin (PRL) is a hormone produced by the pituitary gland with innumerable functions, such as lactation, reproduction,
osmotic and immune regulation. The present work describes the synthesis of hPRL in human embryonic
kidney (HEK293) cells, transiently transfected with the pcDNA-3.4-TOPO® vector carrying the hPRL cDNA. A concentration
of ~ 20 mg/L, including glycosylated (G-hPRL) and non-glycosylated (NG-hPRL) human prolactin, was obtained,
with ~ 19% of G-hPRL, which is higher than that observed in CHO-derived hPRL (~ 10%) and falling within the wide
range of 5–30% reported for pituitary-derived hPRL. N-Glycoprofiling analysis of G-hPRL provided: (i) identification of
each N-glycan structure and relative intensity; (ii) average N-glycan mass; (iii) molecular mass of the whole glycoprotein
and relative carbohydrate mass fraction; (iv) mass fraction of each monosaccharide. The data obtained were compared
to pituitary- and CHO-derived G-hPRL. The whole MM of HEK-derived G-hPRL, determined via MALDI–TOF-MS,
was 25,123 Da, which is 0.88% higher than pit- and 0.61% higher than CHO-derived G-hPRL. The main difference with
the latter was due to sialylation, which was ~ sevenfold lower, but slightly higher than that observed in native G-hPRL.
The “in vitro” bioactivity of HEK-G-hPRL was ~ fourfold lower than that of native G-hPRL, with which it had in common
also the number of N-glycan structures.
Como referenciar
SILVA, FELIPE D.; OLIVEIRA, JOÃO E.; FREIRE, RENAN P.; SUZUKI, MIRIAM F.; SOARES, CARLOS R.; BARTOLINI, PAOLO. Expression of glycosylated human prolactin in HEK293 cells and related N‑glycan composition analysis. AMB Express, v. 9, n. 1, p. 1-11, 2019. DOI: 10.1186/s13568-019-0856-8. Disponível em: http://repositorio.ipen.br/handle/123456789/30430. Acesso em: 30 Dec 2025.
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.