An immunoenzymatic procedure for human apo B-containing particles quantification using monoclonal antibodies

Area of Science:

• Biochemistry
• Immunology
• Clinical Chemistry

Background:

• Apolipoprotein B (apo B) is a key component of atherogenic lipoproteins.
• Accurate quantification of apo B-containing particles is crucial for cardiovascular risk assessment.
• Current methods may have limitations in precision or accessibility.

Purpose of the Study:

• To develop and validate a novel enzyme-linked immunosorbent assay (ELISA) for quantifying apolipoprotein B (apo B) particles.
• To evaluate the performance of different monoclonal antibodies (MAbs) for apo B-100 detection.
• To establish a reliable method for assessing apo B-lipoprotein levels in plasma.

Main Methods:

• Development of a monoclonal antibody-based ELISA.
• Utilized native low-density lipoprotein (LDL) and reference serum for standard curve preparation.
• Examined three laboratory-produced anti-apo B-100 monoclonal antibodies (4A6E3, 6A10B10, 2D9) with varying affinities.

Main Results:

• The monoclonal antibodies exhibited apparent affinity constants in the range of 0.63 x 10^8 to 2.9 x 10^9 M^-1.
• A standard curve was successfully generated for apo B-LDL concentrations from 0.1 to 4.0 microg/ml.
• The developed ELISA allows for the quantification of apo B-containing particles in plasma samples.

Conclusions:

• A new, sensitive ELISA method has been established for quantifying apo B-containing particles.
• The assay utilizes specific monoclonal antibodies against apo B-100.
• This quantification method holds potential for improved assessment of coronary artery disease risk.