Crucian carp (Carassius carassius) VTG monoclonal antibody: Development and application

Abstract

The vitellogenin (VTG) in fish has been used as an important biomarker for monitoring endocrine disrupting compounds (EDCs). This paper reports the development of a new monoclonal antibody (McAb) against the VTG of crucian carp (Carassius carassius). The McAb has a molecular weight of 149.4 kDa (heavy chain: 53.1 kDa; light chain: 21.6 kDa), and double diffusion indicated that it belongs to the IgG1 subclass. The titer is 10 5 -10 6 and the affinity constant (K aff ) is 7.0 Â 10 8 L/mol, showing the high specificity and sensitivity of the antibody. The established sandwich ELISA was sensitive with a detection limit of VTG 0.98 ng/mL. The cross-reactivity of antibody was detected in cyprinids such as rare minnow, zebrafish, and carp. This ELISA was used to detect the variation of VTG in crucian carp exposed to secondary effluent and reclaimed water from the Gaobeidian sewage treatment plant (SRP). The VTG induction in secondary effluent was higher than that in reclaimed effluent and the VTG levels in juvenile crucian carp increased with increasing exposure time. The VTG concentration in male fish from downstream of the Gaobeidian STP was 88.627827.73 mg/mL, while that from a control site was undetectable. r

Evaluation of estrogenicity of sewage effluent and reclaimed water using vitellogenin as a biomarker

“…Blood plasma was assayed for VTG concentration using the previously validated, sandwich enzyme‐linked immunosorbent assay [22]. Briefly, the polyclonal antibodies against crucian carp VTG (c‐VTG) were coated on a 96‐well microtiter plate for a night at 4°C.…”

“…In the present study, VTG analysis was used to evaluate the estrogenic activities of secondary effluent and reclaimed water from the Gaobeidian STP (G‐STP) in Beijing (China). Crucian carp ( Carassius carassius ), a common Chinese species, was exposed to the secondary effluent and reclaimed water treated with the conventional process, and the plasma VTG level was analyzed using an enzyme‐linked immunosorbent assay [22]. For comparison, the estrogenic activities of the secondary effluent and reclaimed water also were evaluated using the recombinant yeast assay.…”

Effects of adrenergic agents on the expression of zebrafish (Danio rerio) vitellogenin Ao1

“…4), which indicates that the regulation of the adrenergic agents on zebrafish vtgAo1 expression is not estrogen signaling dependent. Previous studies have shown that the VTG molecule is highly conserved within the same family, and the cross-reactivity of antibody was detected in cyprinids such as the rare minnow, zebrafish, and carp (Tyler et al, 1996;An et al, 2007). Significant induction of the circulating vtgAo1 level by the MOXI was detected in the plasma with the carp Vtg ELISA Kit, even with the presence of TAM (Fig.…”

Proteomic characterization of vitellogenins from three species of South American fresh water fish

“…Although, some subunits of Vtgs are highly conserved and share a common general structure among fish (Finn, 2007), its complexity due to posttranslational modifications and translocation processes, often results, in poor antigenic cross-reactivity, even between closely related species (Watts et al, 2003;An et al, 2007). This makes a necessary more in-depth study of Vtg for every species of fish.…”