Version 1
: Received: 29 May 2024 / Approved: 30 May 2024 / Online: 30 May 2024 (13:17:46 CEST)
How to cite:
Lima, E. R.; Sevilhano, T.; Caramori, T.; Oliveira, J. E.; Suzuki, M. F.; Torati, L. S.; Bartolini, P.; Peroni, C. N. Physical Chemical, Immunological and Biological Characterization of Arapaima 2 gigas Follicle-Stimulating Hormone (ag-FSH) synthesized in HEK293 Cells. Preprints2024, 2024052027. https://doi.org/10.20944/preprints202405.2027.v1
Lima, E. R.; Sevilhano, T.; Caramori, T.; Oliveira, J. E.; Suzuki, M. F.; Torati, L. S.; Bartolini, P.; Peroni, C. N. Physical Chemical, Immunological and Biological Characterization of Arapaima 2 gigas Follicle-Stimulating Hormone (ag-FSH) synthesized in HEK293 Cells. Preprints 2024, 2024052027. https://doi.org/10.20944/preprints202405.2027.v1
Lima, E. R.; Sevilhano, T.; Caramori, T.; Oliveira, J. E.; Suzuki, M. F.; Torati, L. S.; Bartolini, P.; Peroni, C. N. Physical Chemical, Immunological and Biological Characterization of Arapaima 2 gigas Follicle-Stimulating Hormone (ag-FSH) synthesized in HEK293 Cells. Preprints2024, 2024052027. https://doi.org/10.20944/preprints202405.2027.v1
APA Style
Lima, E. R., Sevilhano, T., Caramori, T., Oliveira, J. E., Suzuki, M. F., Torati, L. S., Bartolini, P., & Peroni, C. N. (2024). Physical Chemical, Immunological and Biological Characterization of Arapaima 2 gigas Follicle-Stimulating Hormone (ag-FSH) synthesized in HEK293 Cells. Preprints. https://doi.org/10.20944/preprints202405.2027.v1
Chicago/Turabian Style
Lima, E. R., Paolo Bartolini and Cibele Nunes Peroni. 2024 "Physical Chemical, Immunological and Biological Characterization of Arapaima 2 gigas Follicle-Stimulating Hormone (ag-FSH) synthesized in HEK293 Cells" Preprints. https://doi.org/10.20944/preprints202405.2027.v1
Abstract
The cDNA sequences of the common gonadotrophic hormone α-subunit (ag-GTHα) and 13 of the specific follicle-stimulating hormone β-subunit (ag-FSHβ) of the giant Amazonian fish Ara-14 paima gigas, have been previously isolated by our research group. A synthesis of ag-FSH in HEK293 15 cells and its purification and preliminary characterization were also carried out. In the present work, 16 ag-FSH was obtained and purified from the same host cells and an extensive physical chemical 17 characterization was performed via RP-HPLC, HPSEC and MALDI-TOF-MS. Ag-FSH, when com-18 pared to human FSH (hFSH), showed a higher hydrophobicity by RP-HPLC and a higher molecular 19 mass (MM) via HPSEC. The same higher MM was also confirmed via MALDI-TOF-MS: 35,353 Da 20 for ag-FSH against 31,969 Da for hFSH. Its immunological activity was also confirmed via an hFSH 21 ELISA, in comparison with the highly purified pituitary preparation of hFSH-AFP7298A, from the 22 National Hormone and Pituitary Program (NHPP-USA). Finally, an in-vitro bioassay, based on the 23 stimulated release of 11-ketotestosterone (11-KT) from immature A. gigas testis, quantified its bio-24 logical activity in comparison with human chorionic gonadotropin (hCG) and with the WHO Inter-25 national Standard of recombinant hFSH (NIBSC-08/282) This suggests that injections of ag-FSH in 26 A. gigas and ag-FSH cDNA Gene Therapy applications should improve the reproductive functions 27 of this threatened species.
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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