A sandwich enzyme-linked immunosorbent assay for beta 2-glycoprotein I

Area of Science:

• Biochemistry
• Immunology
• Clinical Chemistry

Background:

• Beta 2-glycoprotein I (Apolipoprotein H) is a plasma protein with implications in thrombosis and autoimmune diseases.
• Accurate quantification of beta 2-glycoprotein I is crucial for diagnosing and monitoring certain medical conditions.

Purpose of the Study:

• To develop and validate a sensitive solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) for quantifying beta 2-glycoprotein I.
• To assess the assay's performance characteristics in urine samples and its applicability to serum.

Main Methods:

• Development of a solid-phase sandwich ELISA.
• Characterization of assay parameters including working range, detection limit, and precision (within-plate and between-batch CV).
• Evaluation of analyte recovery in spiked urine samples.

Main Results:

• The developed ELISA demonstrates a working concentration range of 5-40 µg/L and a detection limit of approximately 1.4 µg/L.
• High precision was observed with within-plate CVs of 1.4-2.1% and between-batch CVs of 5.2-6.0%.
• Excellent recovery rates (96-110%) indicate assay accuracy for beta 2-glycoprotein I in urine.

Conclusions:

• A robust and sensitive ELISA for beta 2-glycoprotein I determination in urine has been successfully developed.
• The assay exhibits favorable precision and accuracy, making it suitable for clinical applications.
• The method's adaptability for serum analysis broadens its diagnostic potential.