Sandwich immunoassay for determination of vitellogenin in golden grey mullet (Liza aurata) serum as a field exposure biomarker.
Vitellogenin (VTG) is a protein produced in the liver of oviparous animals in response to oestrogens. Abnormal production of VTG by males, therefore, is used as a biological indicator of exposure to xeno-oestrogens. In this study, a sandwich-ELISA for measuring VTG in Liza aurata (golden grey mullet) was developed and validated. Plasma VTG was purified from 17beta-oestradiol-injected immature individuals of mullet, by size-exclusion and ion-exchange chromatography. Polyclonal antibodies against VTG were raised in rabbits. A sensitive immunoassay was developed for measurement of vitellogenin in L. aurata serum, reaching a quantification limit of 0.01 microg mL(-1) and a dynamic range from 0.02 to 2 microg mL(-1). The assay is specific, because high levels (>100 microg mL(-1)) of carp (Cyprinus carpio), goldfish (Carassius auratus), tilapia (Oreochromis niloticus), tench (Tinca tinca), rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and frog (Rana perezi) purified VTG, give negligible responses. The assay was used to analyse plasma samples from wild mullet.
Asturiano
Juan F
JF
Departamento de Ciencia Animal, Universidad Politécnica de Valencia, Camino de Vera s/n, 46071, Valencia, Spain.
Romaguera
Francisco
F
Aragón
Pilar
P
Atienza
Julia
J
Puchades
Rosa
R
Maquieira
Angel
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D004958:Estradiol / Q000008:administration & dosage / Q000633:toxicity; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008297:Male; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D023701:Smegmamorpha / Q000378:metabolism*; D014819:Vitellogenins / Q000097:blood*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D014819:Vitellogenins; D004958:Estradiol