A solid-phase luminescent immunoassay for human chorionic gonadotropin using Pholas dactylus bioluminescence.

Area of Science:

• Biochemistry
• Immunology
• Analytical Chemistry

Background:

• Immunoassays are crucial for detecting biomarkers like human chorionic gonadotropin (hCG).
• Developing sensitive and reliable detection methods is essential for accurate diagnostics.
• Luminescent assays offer potential advantages in sensitivity and detection limits.

Purpose of the Study:

• To develop a solid-phase luminescent immunoassay (LIA) for quantifying human chorionic gonadotropin (hCG).
• To evaluate the sensitivity and performance of the developed LIA compared to existing methods.

Main Methods:

• Development of a luminescent immunoassay using Pholas dactylus luciferin and horseradish peroxidase (HRP).
• Coupling of HRP with anti-hCG antibody for detection.
• Immobilization of hCG antigen in antibody-coated plastic tubes.
• Detection of immunocomplex via light emission.

Main Results:

• The assay detected as little as 0.1 fmol of HRP.
• Light emission showed linearity with hCG concentration in the range of 0.5-100 ng/ml.
• The LIA demonstrated comparable sensitivity (0.5 ng hCG/ml) and good correlation with Radioimmunoassay (RIA).

Conclusions:

• A solid-phase luminescent immunoassay (LIA) for hCG detection has been successfully developed.
• The LIA is sensitive, reliable, and correlates well with RIA, offering a viable alternative for hCG quantification.