LIMA, ELIANA R.SEVILHANO, THAIS C.A.FEITOSA, THAIS C.OLIVEIRA, JOAO E.SUZUKI, MIRIAM F.TORATI, LUCAS S.BARTOLINI, PAOLOPERONI, CIBELE N.2026-02-272026-02-272025LIMA, ELIANA R.; SEVILHANO, THAIS C.A.; FEITOSA, THAIS C.; OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; TORATI, LUCAS S.; BARTOLINI, PAOLO; PERONI, CIBELE N. Extensive characterization of arapaima gigas follicle-stimulating hormone (ag-Fsh) synthesized in HEK293 cells. <b>Fishes</b>, v. 10, n. 12, p. 1-12, 2025. DOI: <a href="https://dx.doi.org/10.3390/fishes10120607">10.3390/fishes10120607</a>. Disponível em: https://repositorio.ipen.br/handle/123456789/49375.2410-3888https://repositorio.ipen.br/handle/123456789/49375The cDNA sequences of the common gonadotrophic hormone alpha-subunit (ag-Gth alpha) and of the specific follicle-stimulating hormone beta-subunit (ag-Fsh beta) of the giant Amazonian fish Arapaima gigas have been previously isolated by our research group. A synthesis of ag-Fsh in HEK293 cells and its purification and preliminary characterization were also carried out. In the present work, ag-Fsh was obtained and purified from the same host cells, and an extensive physical chemical characterization was performed via RP-HPLC, HPSEC, and MALDI-TOF-MS. Ag-Fsh, when compared to human FSH (hFSH), showed a higher hydrophobicity by RP-HPLC and a higher molecular mass (MM) via HPSEC. The same higher MM was also confirmed via MALDI-TOF-MS: 35,353 Da for ag-FSH against 31,969 Da for hFSH. Its immunological activity was also confirmed via an hFSH ELISA, in comparison with the highly purified pituitary preparation of hFSH-AFP7298A, from the National Hormone and Pituitary Program (NHPP-USA), with a clear response that was, however, 1560-fold lower when compared to the purest preparation. Finally, an in vitro bioassay, based on the stimulated release of 11-ketotestosterone (11-KT) from immature A. gigas testis, quantified ag-Fsh biological activity in comparison with human chorionic gonadotropin (hCG) and with human pituitary FSH-AFP7298A, showing a potency clearly higher than that of hCG. This suggests that injections of ag-Fsh in A. gigas and ag-Fsh cDNA gene therapy applications could be used for improving the reproductive functions of this threatened species.1-12engopenAccessArtigo de periódico121010.3390/fishes10120607https://orcid.org/0000-0001-8194-523073.857.00