Validation of a lipoprotein(a) particle concentration assay by quantitative lipoprotein immunofixation electrophoresis

Philip A. Guadagno, Erin G. Summers Bellin, William S. Harris, Thomas D. Dayspring, Daniel M. Hoefner, Dawn L. Thiselton, Brant Stanovick, G. Russell Warnick, Joseph P. McConnell

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


Background: Low-density lipoprotein (LDL) particle (P, or molar) concentration has been shown to be a more sensitive marker of cardiovascular disease (CVD) risk than LDL cholesterol. Although elevated circulating lipoprotein(a) [Lp(a)] cholesterol and mass have been associated with CV risk, no practicable method exists to measure Lp(a)-P. We have developed a method of determining Lp(a)-P suitable for routine clinical use. Methods: Lipoprotein immunofixation electrophoresis (Lipo-IFE) involves rigidly controlled electrophoretic separation of serum lipoproteins, probing with polyclonal apolipoprotein B antibodies, then visualization after staining with a nonspecific protein stain (Acid Violet). Lipo-IFE was compared to the Lp(a) mass assay for 1086 randomly selected patient samples, and for 254 samples stratified by apo(a) isoform size. Results: The Lipo-IFE method was shown to be precise (CV <. 10% above the 50. nmol/l limit of quantitation) and linear across a 16-fold range. Lipo-IFE compared well with the mass-based Lp(a) assay (r= 0.95), but was not affected by variations in apo(a) isoform size. With a throughput of 100 samples in 90. min, the assay is suitable for use in the clinical laboratory. Conclusions: The Lipo-IFE method will allow Lp(a)-P to be readily tested as a CVD risk factor in large-scale clinical trials.

Original languageEnglish (US)
Pages (from-to)219-224
Number of pages6
JournalClinica Chimica Acta
StatePublished - Jan 5 2015
Externally publishedYes


  • Apo(a) isoform size
  • Cholesterol
  • Electrophoresis
  • Immunofixation
  • Lipoprotein particles
  • Lipoprotein(a)

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical


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