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Vitellogenin in black turtle ( Chelonia mydas agassizii ): Purification, partial characterization, and validation of an enzyme‐linked immunosorbent assay for its detection
Author(s) -
SifuentesRomero Itzel,
VázquezBoucard Celia,
SierraBeltrán Arturo P.,
Gardner Susan C.
Publication year - 2006
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1897/05-063r2.1
Subject(s) - polyclonal antibodies , vitellogenin , turtle (robot) , biology , sea turtle , western blot , microbiology and biotechnology , enzyme , antibody , chromatography , chemistry , biochemistry , fishery , gene , genetics
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.