Antibody development to testosterone and its application in capillary electrophoresis-based immunoassay

Area of Science:

• Analytical Chemistry
• Biochemistry
• Forensic Science

Background:

• Accurate testosterone quantification is crucial for clinical diagnostics and doping control.
• Existing immunoassay methods may lack the speed, selectivity, or sensitivity required for certain applications.

Purpose of the Study:

• To develop a rapid, simple, and highly selective testosterone competitive immunoassay.
• To utilize capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection for enhanced sensitivity and specificity.
• To evaluate the method's potential for testosterone doping control in urine samples.

Main Methods:

• A competitive immunoassay format was employed using synthesized fluorescence-labeled testosterone as a tracer.
• Two polyclonal antibodies (Ab(3) and Ab(17)) and their respective tracers were optimized.
• The optimized Ab(3) system was used for testosterone quantification via CE-LIF detection.

Main Results:

• The developed method demonstrated a wide working range of 3.70-2000 ng/mL.
• A low limit of detection (LOD) of 1.11 ng/mL was achieved.
• Testing with normal and positive urine samples confirmed the method's applicability.

Conclusions:

• The CE-LIF based immunoassay offers a rapid, simple, and selective approach for testosterone quantification.
• The method's performance characteristics suggest its utility in testosterone doping control.
• This technique provides a valuable tool for monitoring testosterone levels in biological samples.