Development of a Sensitive Luminometric Immunoassay for Determining Baseline Seasonal Changes in Serum Vitellogenin Levels in Male Flounder (Pleuronectes Yokohamae)

Abstract

Vitellogenin is a sensitive biomarker used to study the effects of artificial estrogens in aquatic environments. We developed and optimized a luminometric immunoassay that was able to detect trace amounts of vitellogenin in the serum of male flounder collected from an uncontaminated reference site. The lowest measurable concentration of vitellogenin in the serum was approximately 0.08 ng/ml with purified protein diluted in buffer. The reproducibility of the vitellogenin measurements was determined by the analysis of triplicate samples and found to be about 5.3%, based on serum samples containing vitellogenin at 2.0 ng/ml. This method was successfully applied to samples collected from an uncontaminated reference site for the monitoring of baseline levels of serum vitellogenin in male flounder.