Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system

Area of Science:

• Analytical Chemistry
• Biotechnology
• Electrochemistry

Background:

• Progesterone (P4) is a crucial steroidal hormone for human and animal health.
• Accurate quantification of P4 is essential for various veterinary and clinical applications.
• Existing methods for P4 detection can be time-consuming or require extensive sample preparation.

Purpose of the Study:

• To develop a rapid and sensitive immunosensor for quantifying progesterone (P4) in bovine serum.
• To integrate an electrochemical immunosensor with a microfluidic system for efficient sample processing.
• To evaluate the performance of the developed immunosensor in terms of sensitivity, detection limits, and applicability.

Main Methods:

• Development of an immunosensor using a glassy carbon (GC) electrode immobilized with anti-P4 antibodies.
• Integration of the immunosensor into a microfluidic system for automated sample handling.
• Utilizing a competitive immunoassay format with horseradish peroxidase (HRP) labeling.
• Electrochemical detection via chronoamperometry measuring HRP enzyme activity.

Main Results:

• The immunosensor demonstrated a linear working range for P4 detection from 0.5 to 12.5 ng/mL.
• Achieved a low detection limit (DL) of 0.2 ng/mL and quantification limit (QL) of 0.5 ng/mL.
• Exhibited higher sensitivity compared to traditional ELISA methods, with comparable detection limits.
• Enabled up to 100 sample determinations without prior sample pre-treatment.

Conclusions:

• The developed microfluidic-integrated electrochemical immunosensor provides a fast, sensitive, and efficient method for P4 quantification in bovine serum.
• This technology is suitable for routine veterinary, clinical, and biological assays requiring P4 level monitoring.
• The immunosensor's ability to handle multiple samples without pre-treatment enhances its practical utility.