Physical chemical, immunological and biological characterization of arapaima 2 gigas follicle-stimulating hormone (ag-FSH) synthesized in HEK293 cells
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The cDNA sequences of the common gonadotrophic hormone α‐subunit (ag‐GTHα) and
of the specific follicle‐stimulating hormone β‐subunit (ag‐FSHβ) 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). Finally, an in‐vitro bioassay,
based on the stimulated release of 11‐ketotestosterone (11‐KT) from immature A. gigas testis,
quantified its biological activity in comparison with human chorionic gonadotropin (hCG) and with
the WHO International Standard of recombinant hFSH (NIBSC‐08/282) This suggests that injections
of ag‐FSH in A. gigas and ag‐FSH cDNA Gene Therapy applications should improve the
reproductive functions of this threatened species.
Como referenciar
LIMA, ELIANA R.; SEVILHANO, THAIS; CARAMORI, THAIS; OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; TORATI, LUCAS S.; BARTOLINI, PAOLO; PERONI, CIBELE N. Physical chemical, immunological and biological characterization of arapaima 2 gigas follicle-stimulating hormone (ag-FSH) synthesized in HEK293 cells. Preprints.org, p. 1-9, 2024. DOI: 10.20944/preprints202405.2027.v1. Disponível em: https://repositorio.ipen.br/handle/123456789/50063. Acesso em: 30 Jun 2026.
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.