Vitellogenin in black turtle (Chelonia mydas agassizii): purification, partial characterization, and validation of an enzyme-linked immunosorbent assay for its detection.
Black turtle plasmatic vitellogenin (VTG) was purified from 17beta-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 microg/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.
Sifuentes-Romero
Itzel
I
Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Mar Bermejo No. 195 Col. Playa Palo de Sta. Rita, CP 23090, La Paz, BCS, Mexico.
Vázquez-Boucard
Celia
C
Sierra-Beltrán
Arturo P
AP
Gardner
Susan C
SC
D000818:Animals; D000906:Antibodies; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D052244:Endocrine Disruptors / Q000506:poisoning; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000502:physiology; D008297:Male; D011817:Rabbits; D012680:Sensitivity and Specificity; D014426:Turtles; D014819:Vitellogenins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*
D000906:Antibodies; D015415:Biomarkers; D052244:Endocrine Disruptors; D014819:Vitellogenins; D004958:Estradiol