An ELISA procedure for human Lp(a) quantitation using monoclonal antibodies.

The present study describes a monoclonal antibody-based enzyme immunoassay (ELISA) for the quantitation of lipoprotein(a), Lp(a), in human plasma. Two antibodies to Lp(a), 2F4E7 and 8G12G7, were produced and characterized as specific and high affinity antibodies against Lp(a). A reference control serum was utilized to prepare the standard curve in a Lp(a) concentration range from 0.015 to 0.5 ug/ml. A biotinylated monoclonal antibody against apoB-LDL was used as the second antibody. The comparison of the standardized ELISA using mAb 2F4E7 with an ELISA using a characterized mAb against Lp(a) (clone KO9) as capture antibody showed that the Lp(a) concentration of two standard sera was similar with both assays. Furthermore, when compared with an electroimmunoassay kit, similar Lp(a) concentrations for the standard were also obtained.

A sandwich ELISA based on anti-apo(a) and anti-apo B monoclonal antibodies for lipoprotein(a) measurement.

Lipoprotein(a) (Lp(a)) is one of the most important independent risk factors for the prediction of premature atherosclerosis. Lp(a) is a low-density lipoprotein (LDL)-like particle which contains a glycoprotein (apoprotein(a)) disulfide linked to apo B-100. We describe a sandwich ELISA based on an anti-apo(a) monoclonal antibody (MAb) and an anti-apo B MAb for the quantitative determination of Lp(a) in human serum. The assay is sensitive, precise and specific. Samples with different apo(a) isoforms had a linear response in a range of 3-70 mg/dl of Lp(a). Correlations between the ELISA and a commercial ELISA, an immunoradiometric assay and electroimmunodiffusion were 0.92, 0.96 and 0.98, respectively. The frequency distribution of Lp(a) concentration in blood donors showed the skew toward the right reported in other populations. Patients with angiographically assessed coronary atherosclerosis had three times higher levels of Lp(a) than those with no signs of coronary atherosclerosis.