Chemiluminescence sequential injection immunoassay for vitellogenin using magnetic microbeads

Area of Science:

• Analytical Chemistry
• Biochemistry
• Environmental Science

Background:

• Vitellogenin (Vg) is a key biomarker for endocrine disruption in fish.
• Accurate and sensitive detection of Vg is crucial for environmental monitoring.
• Existing methods for Vg determination can be time-consuming or lack sensitivity.

Purpose of the Study:

• To develop a rapid and sensitive immunoassay for carp vitellogenin (Vg).
• To utilize sequential injection analysis (SIA) with magnetic beads for Vg determination.
• To establish an efficient method for assessing fish health and environmental quality.

Main Methods:

• Development of a sandwich immunoassay using anti-Vg monoclonal antibody immobilized on magnetic beads.
• Sequential injection analysis (SIA) system with a chemiluminescence detector.
• Magnetic bead manipulation via a samarium-cobalt magnet for trapping and release in a flow cell.
• Chemiluminescence detection based on horseradish peroxidase (HRP) activity.

Main Results:

• The immunoassay demonstrated high sensitivity with a lower detection limit of approximately 2 ng/mL for carp Vg.
• Optimal incubation times for both immunoreactions were determined to be 20 minutes.
• A concave calibration curve was obtained for Vg concentrations ranging from 2-100 ng/mL.

Conclusions:

• The developed SIA-based immunoassay offers a rapid and sensitive method for carp Vg determination.
• This technique is suitable for monitoring endocrine disruption in fish populations.
• The use of magnetic beads simplifies sample handling and enhances assay efficiency.