Identification, partial characterization, and use of grey mullet (Mugil cephalus) vitellogenins for the development of ELISA and biosensor immunoassays

Abstract

Vitellogenin (Vtg) has proven to be a sensitive and simple biomarker in determining sex, sexual maturity, and xenoestrogenic effects in fish. Thus, our investigation has been focused on identification, partial characterization, and quantification of grey mullet (Mugil cephalus) Vtg through the use of a variety of biochemical and immunological analytical techniques. Mullet is considered both a promising aquaculture candidate and an important species for improving sediment quality in polyculture systems. In the first part of this work, grey mullet Vtg was purified from plasma of 17b-estradiol (E2)-induced male fish by a one-step chromatographic protocol, and partially characterized. Specific polyclonal antibodies were then raised against the mullet Vtg, and both an indirect ELISA and an optical immunosensor were set up and validated to quantify plasma Vtg. The indirect ELISA and the optical immunosensor assay developed showed linear measuring in the range 56.8-1047.1 ng mL-1 and 70-739 ng mL-1 Vtg concentrations in standard solutions, respectively. The results obtained suggest that the indirect ELISA allows Vtg detection over a wide dynamic range, thus resulting more suitable for rapid and sensitive sample screening. Therefore, we suggest that the direct immunosensor is a promising tool which needs more investigation to improve the sensitivity.

An In Silico and In Vitro Study for Investigating Estrogenic Endocrine Effects of Emerging Persistent Pollutants Using Primary Hepatocytes from Grey Mullet (Mugil cephalus)

“…VTG concentrations in the culture medium of Mugil cephalus hepatocytes were determined using an ELISA method previously published [25]. Cell culture media were diluted 1:8 as reported for routinely diluted media samples by Navas and Segner [49].…”

“…Activation of the ER-mediated signaling pathway has been extensively studied in several models, particularly fish in which feminization has been considered a direct result of xenoestrogen contamination [19][20][21][22][23]. In this regard, the ER-induced hepatic vitellogenin (Vtg) production is typically used to confirm exposure to estrogenic compounds in male fish [24,25]. Of the different fish organ cells, liver cells are widely used in in vitro primary culture models due to their ability to retain native liver properties including estrogen responsiveness [26][27][28].…”

An Ultrasensitive Photoluminescent Immunosensing of Lipovitellin by Using N-doped Graphene Quantum Dots

“…Immunosensing strategy is the most widely used method in biomolecular detection. At present, the main detection methods for Lv including in immune diffusion method [16], chemiluminescence method [17], enzymelinked immunoassay [18], surface enhanced Raman scattering (SERS) [19] and optical waveguide lightmode spectroscopic immunosensing [20,21]. These methods have good sensitivities and specificities.…”

Ultrasensitive Fluorescent “ON-OFF” Label-Free Immunosensor for Detection of Vitellogenin of Marine Medaka

“…In recent years, some immunoassay techniques, such as enzyme-linked immunosorbent assay (ELISA) [19][20][21], electrochemical sensors [22,23], and optical sensors [24][25][26], were used to test Vtg. Yi et al [19] used an ELISA method for the determination of Vtg in Marine medlifish with a detection limit of 3.1 ng/mL and a working range of 15.6-500 ng/mL.…”