title: ELISA for human serum leucine-rich alpha-2-glycoprotein-1 employing cytochrome c as the capturing ligand.
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 microg/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.
authors.affiliation: Department of Microbiology and Center for Immunology, University of Minnesota, MN 55455, United States.
authors.forename: Starchild
authors.initials: S
authors.lastname: Weivoda
authors.forename: John D
authors.initials: JD
authors.lastname: Andersen
authors.forename: Aunica
authors.initials: A
authors.lastname: Skogen
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Schlievert
authors.forename: Donna
authors.initials: D
authors.lastname: Fontana
authors.forename: Timothy
authors.initials: T
authors.lastname: Schacker
authors.forename: Paul
authors.initials: P
authors.lastname: Tuite
authors.forename: Janet M
authors.initials: JM
authors.lastname: Dubinsky
authors.forename: Ronald
authors.initials: R
authors.lastname: Jemmerson
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry*; D015415:Biomarkers / Q000097:blood; D015153:Blotting, Western; D002097:C-Reactive Protein / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D002846:Chromatography, Affinity; D045304:Cytochromes c / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D006023:Glycoproteins; C015246:LRG1 protein, human; D002097:C-Reactive Protein; D045304:Cytochromes c