Vitellogenin in black turtle (Chelonia mydas agassizii): Purification, partial characterization, and validation of an enzyme-linked immunosorbent assay for its detection

• Published in: Environmental toxicology and chemistry 2006-02, Vol.25 (2), p.477-485
• Main Authors: Sifuentes-Romero, Itzel, Vázquez-Boucard, Celia, Sierra-Beltrán, Arturo P., Gardner, Susan C.
• Format: Article
• Language: English
• Subjects: Amino acids; Animal, plant and microbial ecology; Animals; Antibodies; Applied ecology; Aquatic reptiles; Assaying; Assessments; Biological and medical sciences; Biomarker; Biomarkers - analysis; Biomarkers - chemistry; C (programming language); Caretta caretta; Chelonia mydas agassizii; Chromatography; Coefficient of variation; Copyrights; Division; Documents; Ecotoxicology, biological effects of pollution; Endocrine Disruptors - poisoning; Enzyme-Linked Immunosorbent Assay; Enzymes; Eretmochelys imbricata; Estradiol - physiology; Fundamental and applied biological sciences. Psychology; General aspects; Health; Homology; Human influences; Immunoassay; Male; Males; Monomers; Presses; Purification; Rabbits; Reptiles & amphibians; Sea turtles; Sensitivity and Specificity; Turtles; Vertebrates; Vitellogenin; Vitellogenins - analysis; Vitellogenins - chemistry; Vitellogenins - isolation & purification
• ISSN: 0730-7268; 1552-8618
• DOI: 10.1897/05-063R2.1

Black turtle plasmatic vitellogenin (VTG) was purified fom 177β‐estradiol–induced males using ion‐exchange chromatography. The isolated protein was identified as VTG by its glycolipoprotein nature and amino acid sequence homology with other vertebrate VTG. It was characterized as a 500‐kDa dimer composed of two identical, 200‐ to 240‐kDa monomers. Polyclonal antibodies raised against black turtle VTG showed high titer and specificity, as demonstrated by enzyme‐linked immunosorbent assay and Western blot analysis, respectively. The range of the assay was estimated to be between 15 ng/ml and 2 μg/ml, and the inter‐ and intra‐assay coefficients of variation were 9.4 and 7.3%, respectively. Black turtle antibody cross‐reacted with VTG of two other sea turtle species, Caretta caretta (loggerhead) and Eretmochelys imbricata (hawksbill), extending the applicability of the assay as part of a sea turtle health assessment program.