ELISA for human serum leucine-rich alpha-2-glycoprotein-1 employing cytochrome c as the capturing ligand

Starchild Weivoda, John D. Andersen, Aunica Skogen, Patrick M. Schlievert, Donna Fontana, Timothy W Schacker, Paul J Tuite, Janet M Dubinsky, Ronald Jemmerson

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

Leucine-rich alpha-2-glycoprotein-1 (LRG) is a serum glycoprotein of unknown function that has shown promise based on qualitative assessments as a biomarker for certain diseases including microbial infections and cancer. However, the lack of a quantitative assay for LRG has limited its application. Here an indirect enzyme-linked immunosorbent assay (ELISA) for quantifying LRG in human serum is described in which cytochrome c is employed as the capturing ligand and a monoclonal antibody specific for LRG is used to detect the captured glycoprotein. Application of this assay in quantifying LRG in various patients' sera is demonstrated. The concentration of LRG in sera of control subjects as determined by this assay is approximately 50 μg/ml. Consistent with expectations from published reports, LRG was found to be significantly elevated in the sera of some patients with a bacterial infection (toxic shock syndrome, TSS). LRG was only slightly elevated in patients infected with the human immunodeficiency virus as compared to uninfected control subjects, while normal levels of LRG were observed in patients with non-infectious diseases (inflammatory arthritis and neurological disorders, primarily Parkinson's disease). Although LRG and C-reactive protein (CRP) are both produced by the liver and are classified as acute-phase proteins, there was no significant correlation between the levels of LRG and CRP in the sera of the patients. Thus, LRG and CRP measurements are non-redundant and indicate different physiological contexts. The ELISA described in this report should be useful to further assess serum LRG as a biomarker for clinical diagnostics.

Original languageEnglish (US)
Pages (from-to)22-29
Number of pages8
JournalJournal of Immunological Methods
Volume336
Issue number1
DOIs
StatePublished - Jul 20 2008

Keywords

  • Biomarker
  • Cytochrome c
  • Leucine-rich alpha-2-glycoprotein-1
  • Sandwich ELISA
  • Toxic shock syndrome

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