Establishment of specific enzyme-linked immunosorbent assay (ELISA) for measuring Fsh and Lh levels in medaka (Oryzias latipes), using recombinant gonadotropins

Susann Burow, Romain Fontaine, Kristine von Krogh, Ian Mayer, Rasoul Nourizadeh-Lillabadi, Lian Hollander-Cohen, Yaron Cohen, Michal Shpilman, Berta Levavi-Sivan, Finn Arne Weltzien*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

The paucity of information on understanding the regulatory mechanisms that are involved in the control of piscine Fsh and Lh synthesis, secretion, and function, prompted the present work. Part of the problem is related to the molecular heterogeneity and the unavailability of Fsh and Lh assays for quantifying gonadotropins, in particular assays regarding the measurement of Fsh, and such assays are available today for only a few teleost species. The present study reports the development and validation of competitive ELISAs for quantitative determination of medaka Fsh and Lh by first producing medaka recombinant (md) gonadotropins mdFshβ, mdLhβ, mdFshβα, and mdLhβα by Pichia pastoris, generating specific antibodies against their respective β subunits, and their use within the development of ELISAs. The advantages of this protocol include: • The reproducibility of the ELISA demonstrated was relatively high, as shown by reasonably low intra- (Fsh 2.7%, Lh 3%) and interassay CVs (Fsh 5.3%, Lh 5.7%). • The high degree of parallelism between serial dilutions of the recombinant and native pituitary-derived Fsh and Lh, may be a sign of similar structures and immunologically similarity. • Two new competitive ELISAs for the quantification of medaka Fsh and Lh were established for the first time.

Original languageAmerican English
Pages (from-to)1473-1479
Number of pages7
JournalMethodsX
Volume6
DOIs
StatePublished - 2019

Bibliographical note

Publisher Copyright:
© 2019 The Author(s)

Keywords

  • Competitive enzyme-linked immunosorbent assay
  • Follicle-stimulating hormone
  • Gonadotropin quantification
  • Luteinizing hormone
  • Oryzias latipes
  • Specific antibodies
  • Specific enzyme-linked immunosorbent assay (ELISA) for determining Fsh and Lh levels in medaka

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