Standardization of Lp(a) measurements.
Two direct binding ELISAs were developed in our laboratory for measuring Lp(a) in human plasma. The first ELISA was performed by using a monoclonal antibody to apo(a) bound to the solid phase and a second monoclonal antibody to apo(a) as detecting antibody. The second ELISA was performed by using the same monoclonal antibody bound to the solid phase and an anti-apo B polyclonal antibody as detecting antibody. The immunoreactivity of Lp(a) particles of different size, isolated from subjects with F, B or S2 isoform, were evaluated by the two ELISA methods. Superimposable standard curves were obtained by the three Lp(a) preparations when using the apo(a) detection system, but the Lp(a) containing the largest apo(a) isoform S2 had a significantly reduced slope by the apo B detection method. Lp(a) concentration was determined in plasma samples by the two ELISA methods. When Lp(a) with apo(a) isoform S2 was used to calibrate the assays, similar Lp(a) values were obtained by the two different detecting systems on samples from subjects with isoforms S2, S3 or S4, while values in the samples with B and S1 isoforms were significantly higher. When Lp(a) with isoform B was used as calibrator, comparable Lp(a) values were obtained by the two methods on samples with B isoform, while the values were lower in the samples with the higher molecular weight isoforms when measured by the apo B detection method. A pilot study was conducted to evaluate Lp(a) values obtained by different methods calibrated with a common fresh-frozen serum with a defined apo(a) isoform.(ABSTRACT TRUNCATED AT 250 WORDS)
Albers
J J
JJ
Department of Medicine, University of Washington, Seattle 98103.
Marcovina
S M
SM
D000911:Antibodies, Monoclonal; D001053:Apolipoproteins / Q000276:immunology / Q000592:standards; D001055:Apolipoproteins B / Q000276:immunology / Q000592:standards; D053241:Apoprotein(a); D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D005069:Evaluation Studies as Topic; D006801:Humans; D007120:Immunochemistry; D017270:Lipoprotein(a) / Q000097:blood* / Q000276:immunology / Q000592:standards*; D010316:Particle Size; D012015:Reference Standards
D000911:Antibodies, Monoclonal; D001053:Apolipoproteins; D001055:Apolipoproteins B; D017270:Lipoprotein(a); D053241:Apoprotein(a)