PMID: 16519293Mar 8, 2006Paper

Plasma sampling and freezing procedures influence vitellogenin measurements by enzyme-linked immunoassay in the fathead minnow (Pimephales promelas)

Environmental Toxicology and Chemistry
Julie C BrodeurGary M Klecka

Abstract

The present study compared three different methods for measuring plasma vitellogenin (VTG) in fathead minnow (FHM; Pimephales promelas): A procedure using liquid chromatography with electrospray ionization combined with tandem mass spectrometry (LC/ESI-MS/MS), and two commercial enzyme-linked immunoassay (ELISA) kits using either anti-carp or anti-FHM antibodies. The influence on plasma VTG measurements of using the protease-inhibitor aprotinin during blood sampling and of submitting the plasma samples to a freeze-thaw cycle before analysis also was evaluated. The addition of aprotinin to the blood during sampling significantly reduced the plasma VTG concentrations measured by ELISA, whereas the VTG values measured after plasma samples were submitted to a freeze-thaw cycle were significantly higher than those measured before freezing. This inflating effect of freezing on VTG measurements made by ELISA could be prevented if plasma samples were frozen diluted in citrate buffer containing 16 mg/ml of polyethylene glycol (PEG). In contrast, measurements of VTG made by LC/ESI-MS/MS were unaffected by freezing and, conceptually, are independent from enzymatic degradation. Although the use of aprotinin and PEG effectively reduced the ...Continue Reading

Citations

Jan 3, 2014·Birth Defects Research. Part B, Developmental and Reproductive Toxicology·Katherine Kemler CoadyTroy Alan Marino
Mar 17, 2016·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·Jun WangShaoguo Ru
Apr 19, 2006·Ecotoxicology and Environmental Safety·Kathleen M Jensen, Gerald T Ankley

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