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Enzyme-linked immunoassay for Lp[a].

G M Fless¹, M L Snyder, A M Scanu

¹Department of Medicine, Pritzker School of Medicine, University of Chicago, IL 60637.

Journal of Lipid Research

|May 1, 1989

Summary

A new enzyme-linked immunosorbent assay (ELISA) accurately measures lipoprotein(a) [Lp(a)] in plasma, unaffected by plasminogen. This assay accounts for Lp(a) size variations, crucial for accurate clinical assessment.

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Area of Science:

Biochemistry
Immunology
Clinical Chemistry

Background:

Rabbit antisera against human lipoprotein(a) [Lp(a)] or apolipoprotein(a) [apo(a)] can cross-react with plasminogen.
This cross-reactivity poses challenges for accurate Lp(a) quantification in plasma.
Existing assays may be influenced by plasminogen, affecting Lp(a) measurement reliability.

Purpose of the Study:

To develop a novel assay for quantifying plasma Lp(a) that is insensitive to plasminogen.
To utilize a "sandwich" enzyme-linked immunosorbent assay (ELISA) strategy for improved specificity.
To investigate the impact of Lp(a) heterogeneity on assay performance and population distribution.

Main Methods:

Developed a "sandwich" ELISA using anti-apo(a) as the capture antibody.
Quantified bound Lp(a) (apoB-100-apo(a) complex) using an anti-apoB antibody.
Evaluated assay performance with purified Lp(a) and whole plasma, assessing dose-response curves and reactivity across different apo(a) isoforms.

Main Results:

The developed ELISA is insensitive to plasminogen, ensuring specific Lp(a) measurement.
All tested Lp(a) particles showed parallel dose-response curves and immunological equivalence.
Lp(a) particles with larger apo(a) isoforms exhibited lower reactivity on a weight basis but equivalent reactivity on a molar basis.

Conclusions:

Successfully developed a plasminogen-insensitive ELISA for Lp(a) protein quantification.
Lp(a) and apo(a) heterogeneity significantly impacts assay variation, highlighting the need for appropriate standardization.
Population studies revealed skewed Lp(a) distribution, with all subjects having measurable levels and a mean of 3.2 mg/dl.

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