IgY antibodies, also called egg yolk immunoglobulins, are the only immunoglobulins in egg yolk and transferred in the female from serum to egg yolk to confer passive immunity to embryos and neonates. Using hens instead of mammals as the immunization host brings a number of advantages: Eggs are cheap and readily available; antibody levels in yolks are high; IgY isolation is fast and simple; and what is more; IgY neither binds the rheumatoid factor nor reacts to the mammalian complement factor. All these differences make IgY technology more widely applicable, such as in the production of polyclonal antibodies against various antigens, immunodiagnostics and immunotherapy, and many medical areas in both animals and human. IgY also has a good prospect in human immunocontraception.
Animals ; Chickens ; Egg Yolk ; immunology ; Female ; Humans ; Immunoglobulins ; isolation & purification ; therapeutic use ; Vitellogenins ; immunology

OBJECTIVETo assess the single and combined effects of estrone (E1) and 17β-estradiol (E2) on goldfish (Carassius auratus).

METHODSBatch tests were conducted. Serum levels of vitellogenin (VTG) and E2, gonadosomatic indices (GSI), gonadal DNA damage and liver 7-ethoxyresorufin-O-deethylase (EROD) activity were measured after exposure for 14 days.

RESULTSThe VTG level increased significantly in a concentration-dependent manner. The serum E2 level was significantly higher and the GSI level was significantly lower in goldfish after exposed to the 3 drugs. DNA damage occurred in treated samples and EROD activity was significantly suppressed 7 days after exposure. The joint effect of E1 and E2 was additive with regard to VTG induction.

CONCLUSIONThe results of our study highlight a series of effects of steroidal estrogens on goldfish. Further study is needed to confirm their effect as a whole.

Animals ; Cytochrome P-450 CYP1A1 ; metabolism ; DNA Damage ; drug effects ; Drug Combinations ; Estradiol ; pharmacology ; Estrone ; pharmacology ; Goldfish ; Gonads ; drug effects ; metabolism ; Male ; Vitellogenins ; blood

We developed a new method for soluble expression of phage-displayed scFv antibody specific for zebrafish vitellogenin. The scFv antibody F5 could bind zebrafish vitellogenin specifically in phage-displayed form but not soluble form. The gene of scFv antibody F5 was cloned into vector pET 32a and transferred into Escherichia coli ori DE3. With inducible expression, soluble scFv antibody 32a-F5 was obtained successfully and could also specifically bind to zebrafish vitellogenin. The insoluble expression of phage-displayed scFv antibody was a common problem in the practical use of phage display. This study offered a feasible way to express soluble scFv antibodies with biological activity.
Amino Acid Sequence ; Animals ; Antibody Specificity ; Base Sequence ; Cell Surface Display Techniques ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Single-Chain Antibodies ; biosynthesis ; genetics ; immunology ; Solubility ; Vitellogenins ; immunology ; Zebrafish ; immunology ; metabolism

OBJECTIVES: Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. METHODS: Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. RESULTS: The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was 99.5+/-5.5%. CONCLUSIONS: A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.
Antibodies ; Antibodies, Monoclonal ; Blotting, Western ; Cyprinidae* ; Ecosystem ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Endocrine Disruptors ; Enzyme-Linked Immunosorbent Assay* ; Estradiol ; Estrogens ; Female ; Food Chain ; Humans ; Liver ; Male ; Mass Screening ; Methods ; Molecular Weight ; Oogenesis ; Ovary ; Ovum ; Plasma ; Platypus* ; Sensitivity and Specificity ; Sodium ; Vitellins* ; Vitellogenins* ; Weights and Measures