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

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 x 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 88.62+/-827.73 microg/mL, while that from a control site was undetectable.