The purpose of this study was to develop a direct sandwich enzyme-linked immunosorbent assay (ELISA) for vitellogenin-derived yolk proteins (VDYP) in muscle tissue of pelagic Atlantic blue marlin. Gravid ovaries were extracted with physiologic buffer, and purification steps by chromatography were monitored with a cross-reactive mouse monoclonal antibody produced to swordfish (Xiphias gladius Linnaeus, 1758) VDYP. A polyclonal goat antiserum was produced to the enriched VDYP from which specific IgG was purified by affinity chromatography, using immobilized VDYP as the immunoadsorbent. A direct sandwich ELISA was developed using the purified IgG for both capture and detection of the VDYP in standard samples up to a concentration of 1000 ng/ml, and in physiologic buffer-extracted muscle tissue of test samples, using biotin-streptavidin- peroxidase technology. The assay distinguished vitellogenic from non-vitellogenic females and males in extracts of muscle tissue from 27 specimens collected from different areas of the western North Atlantic Ocean during May-September, 2002-2003. The assay for VDYP, and a second for the fish androgen 11-ketotestosterone are in use to indicate gender of electronically-tagged blue marlin from biopsy samples taken just before fish release.
|Original language||English (US)|
|Number of pages||11|
|Journal||Bulletin of Marine Science|
|State||Published - Mar 1 2006|
ASJC Scopus subject areas
- Aquatic Science