Re-evalution of the measurement of haptoglobin in human plasma samples.

Maria Kløjgaard Skytthe, Anna Lahn Sørensen, Dorle Hennig, Maria Boysen Sandberg, Lars Melholt Rasmussen, Holger Jon Møller, Karsten Skjødt, Jonas Heilskov Graversen, Søren Kragh Moestrup

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Haptoglobin (Hp) is an abundant plasma protein scavenging hemoglobin (Hb) via CD163 on macrophages. This process consumes Hp, which therefore negatively correlates to hemolysis. However, exact measurements of Hp plasma levels are complicated by different phenotypes (Hp1-1, Hp2-1, and Hp2-2) forming different oligomeric states with differences in immunoreactivity. In addition, humans have an immune-cross-reactive Hp-related protein. In the present study, we developed Hp-specific monoclonal antibodies for an accurate Hp analysis of the different Hp phenotypes in a panel of 112 anonymous samples from hospitalized individuals subjected to routine Hp immunoturbidimetric measurements. The data revealed immunoturbidimetry as a reliable method in most cases but also that the use of non-phenotype-specific calibrators leads to substantial bias in the measurement of the Hp-concentration, non at least in Hp1-1 individuals. Furthermore, analysis of the Hb-dependence of the CD163 interaction with Hp1-1 and Hp2-2 showed that a higher ‘cost-effectiveness’ in the consumption of dimeric Hp1-1 versus multimeric Hp phenotypes is a likely contribution to the observed differences in the plasma levels of the Hp phenotypes. In conclusion, the determination of Hp phenotype and the use of phenotype-specific calibrators are essential to obtain a precise estimate of the Hp level in healthy and diseased individuals.

Original languageEnglish
JournalScandinavian Journal of Clinical and Laboratory Investigation
Volume82
Issue number6
Pages (from-to)467-473
ISSN0036-5513
DOIs
Publication statusPublished - Sep 2022

Keywords

  • Haptoglobins
  • blood proteins
  • enzyme-linked immunosorbent assay
  • hemoglobins
  • immunoassay
  • phenotype
  • Phenotype
  • Haptoglobins/genetics
  • Humans
  • Hemoglobins/metabolism
  • Chromosomal Proteins, Non-Histone/genetics
  • Antibodies, Monoclonal

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