Establishment of a novel lectin-antibody ELISA system to determine core-fucosylated haptoglobin

Clin Chim Acta. 2015 Jun 15:446:30-6. doi: 10.1016/j.cca.2015.03.037. Epub 2015 Apr 7.

Abstract

Background: Fucosylated haptoglobin (Fuc-Hpt) is a novel cancer biomarker that increases in various pathological conditions. We previously established a Fuc-Hpt lectin-antibody assay using Aleuria aurantia lectin (AAL), and applied this to diagnose several diseases, including various cancers. AAL recognizes both α1-3/1-4 and α1-6 fucosylation on N/O-linked glycans. These fucosylation types differ in biological function, and in regulation by different fucosyltransferases. Recently, we identified a novel lectin, Pholiota squarrosa lectin (PhoSL), which specifically recognizes α1-6 fucosylation (core-fucosylation).

Methods: We developed a lectin-antibody ELISA kit using PhoSL to determine core-Fuc-Hpt levels in sera from colorectal or pancreatic cancer patients.

Results: Serum levels of AAL-reactive Hpt are higher in pancreatic cancer patients, whereas those of PhoSL-reactive Hpt are higher in colorectal cancer patients. Mass spectrometry analyses of Hpt fucosylation levels were consistent with lectin-antibody ELISA results. Hpt-transfected colorectal cancer cell lines produced significant amounts of core-Fuc-Hpt, suggesting that colorectal cancer tissues produce core-Fuc-Hpt.

Conclusions: These differences in Fuc-Hpt patterns might depend on cancer cells and the surrounding cells, which produce Hpt.

Keywords: AAL; Core fucose; Fucosylation; Haptoglobin; Lectin–antibody ELISA; PhoSL.

MeSH terms

  • Animals
  • Antibodies, Neoplasm / blood*
  • Biomarkers, Tumor / blood*
  • Cells, Cultured
  • Colorectal Neoplasms / blood*
  • Colorectal Neoplasms / diagnosis
  • Enzyme-Linked Immunosorbent Assay / methods
  • Fucosyltransferases / blood
  • Haptoglobins / metabolism*
  • Humans
  • Lectins / blood*
  • Mice
  • Mice, Knockout
  • Pancreatic Neoplasms / blood*
  • Pancreatic Neoplasms / diagnosis
  • Spectrometry, Mass, Electrospray Ionization / methods

Substances

  • Antibodies, Neoplasm
  • Biomarkers, Tumor
  • HP protein, human
  • Haptoglobins
  • Lectins
  • lectin, Aleuria aurantia
  • Fucosyltransferases
  • Glycoprotein 6-alpha-L-fucosyltransferase