Carp vitellogenin detection by an optical waveguide lightmode spectroscopy biosensor

Abstract

A label-free carp vitellogenin sensor has a strong potential for on-site monitoring on the possible contamination of edible fish with endocrine disruptors as a sum parameter in an inland carp farm. In this study, we performed a sensitive detection for carp vitellogenin with a direct-binding optical waveguide lightmode spectroscopy-based immunosensor. Carp vitellogenin bound over the sensor surface quite specifically, judging from the sensor responses according to stepwise antibody immobilization. This was also supported by a negligible sensor response found at bovine serum albumin immobilization. When plotted in double-logarithmic scale for carp vitellogenin concentrations of 0.00675–67.5 nM, a linear relationship was found between analyte concentration and sensor response, together with the limit of detection of 0.00675 nM. The reusability of the immunosensor after the regeneration with 10 mM HCl was reasonably good, as presumed from the coefficient of variability of 6.02% for nine repetitive measurements. The model sample prepared by spiking a purified carp vitellogenin into a 10-fold diluted vitellogenin-free carp serum in 9.45 nM showed the response ratio of 96.70% against 9.45 nM of the purified carp vitellogenin. When a female and male carp sera induced with 17β-estradiol injection were analyzed, biomarker induction was even identifiable at 2000-fold serum dilution.