Micro competition enzyme linked immunosorbant assay for human apolipoprotein B

Area of Science:

• Biochemistry
• Immunology
• Clinical Chemistry

Background:

• Apolipoprotein B (Apo B) is a key component of lipoproteins and a significant risk factor for cardiovascular disease.
• Accurate quantification of Apo B in human plasma is crucial for assessing cardiovascular risk.
• Existing methods for Apo B measurement have limitations, including complexity, cost, and use of radioisotopes.

Purpose of the Study:

• To develop and validate a novel solid phase micro competition enzyme linked immunosorbent assay (microCELIA) for the quantification of apolipoprotein B (Apo B) in human plasma.
• To evaluate the performance characteristics of the developed microCELIA assay.
• To compare the microCELIA assay with established methods like radioimmunoassay (RIA) and electroimmunoassay.

Main Methods:

• Development of a solid phase microCELIA assay where soluble Apo B competes with solid-phase antigen for antibody binding.
• Utilized alkaline phosphatase-labeled goat anti-rabbit IgG antibody for detection.
• Assayed human plasma samples and compared results with RIA and electroimmunoassay reference methods.

Main Results:

• The microCELIA assay demonstrated a minimal quantifiable concentration of 1 mg/L for Apo B.
• Excellent correlation was observed between microCELIA and reference methods: r = 0.97 with RIA and r = 0.92 with electroimmunoassay.
• The microCELIA assay showed less overestimation of hyperlipemic samples compared to electroimmunoassay.

Conclusions:

• The developed microCELIA is a sensitive, simple, and cost-effective method for measuring Apo B in human plasma.
• This enzyme immunoassay (EIA) offers significant advantages, including reduced incubation time and elimination of radioisotopes.
• MicroCELIA provides a reliable alternative to existing techniques for Apo B quantification, aiding in cardiovascular risk assessment.