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Published on: October 7, 2009
Time-resolved immunofluorometric assay for the quantification of lipoprotein(a) in serum
1Institut für Radiologie, Medizinischen Universität zu Lübeck, Germany.
Insights
This study introduces a rapid and robust assay for quantifying lipoprotein(a) (Lp(a)) serum levels. This method aids in evaluating coronary heart disease risk by measuring Lp(a) concentrations.
Area of Science:
- Clinical Chemistry
- Immunology
- Cardiovascular Disease Research
Background:
- Lipoprotein(a) (Lp(a)) is frequently measured to assess coronary heart disease risk.
- While not consistently an independent risk factor for early atherosclerosis, Lp(a) levels are clinically relevant.
- Accurate quantification of Lp(a) is crucial for risk stratification.
Purpose of the Study:
- To describe a time-resolved immunofluorometric assay (TRIFMA) for quantifying human serum Lp(a) levels.
- To establish a rapid, robust, and simple method for Lp(a) measurement.
- To validate the assay's performance characteristics and compare it with existing methods.
Main Methods:
- A two-site immunometric assay utilizing microtitre plates coated with anti-Lp(a) antibodies.
- Biotin-labeled antibodies detected using europium-labeled streptavidin and a DELFIA 1232 fluorometer.
- Assay validation included measuring range, imprecision, interference, and correlation studies.
Main Results:
- The TRIFMA demonstrated a measuring range of 2-1600 mg/l with intra-assay imprecision < 7% and inter-assay imprecision < 12%.
- No interference was observed with plasminogen concentrations up to 2.2 g/l.
- Correlations with enzyme immunoassay (r=0.95) and electroimmunodiffusion (r=0.85) were acceptable, and the assay detected different Lp(a) isoforms.
Conclusions:
- The developed TRIFMA is a reliable and efficient method for quantifying Lp(a) in human serum.
- This assay provides a valuable tool for clinical laboratories assessing cardiovascular risk.
- The assay's ability to detect various Lp(a) isoforms enhances its clinical utility.
Abstract:
Although two recent studies have failed to reveal lipoprotein(a) (LP(a)) serum concentrations > 300 mg/l to be an independent risk factor for early onset of atherosclerosis, Lp(a) serum concentrations are frequently measured to evaluate the additional risk of coronary heart disease. We describe a time-resolved immunofluorometric assay (TRIFMA) for quantifying Lp(a) levels in humans serum using commercially available reagents, which is rapid, robust and simple to perform. The two-site immunometric assay was based on microtitre plates as solid phase coated with a polycloncal anti Lp(a) antibody. The liquid-phase antibody was labelled with biotin and detected by europium labelled streptavidin in the DELFIA 1232 fluorometer. The measuring range was 2-1600 mg/l. The intra-assay imprecision was < 7% (CV), the inter-assay imprecision < 12% (CV). No interference was detected with plasminogen concentration up to 2.2 g/l. There was an acceptable correlation with a commercially available enzyme immunoassay (r = 0.95) and with electroimmunodiffusion (r = 0.85) on 100 routine serum samples measured. The assay appeared to detect different Lp(a) isoforms as dilution curves were parallel for B/F, S2 and S4 isoforms.
