Development of a Novel Enzyme-Linked Immunosorbent Assay Targeting a Neo-Epitope Generated by Cathepsin-Mediated Turnover of Type III Collagen and Its Application in Chronic Obstructive Pulmonary Disease

PLoS One. 2017 Jan 11;12(1):e0170023. doi: 10.1371/journal.pone.0170023. eCollection 2017.

Abstract

A high level of extracellular matrix (ECM) turnover characterizes several lung diseases with fibrotic features. Type III collagen is one of the most abundant collagens in lung parenchyma, and cathepsins play a role in lung pathology, being responsible for tissue remodeling. In this study, we explore the diagnostic features of neo-epitope fragments of type III collagen generated by cathepsins that could reflect the pathological tissue turnover in patients with different diseases. A novel enzyme-linked immunosorbent assay (ELISA) measuring cathepsins B, L, S and K -generated type III collagen fragments (C3C) was developed for assessment in serum and plasma. The assay was biologically validated in serum from patients with chronic obstructive pulmonary disease (COPD). Serological levels of C3C were significantly elevated in patients with COPD compared to healthy controls (p = 0.0006). Levels of C3C in serum and heparin plasma of COPD patients had a highly significant correlation (R2 = 0.86, p<0.0001). The data suggests that the C3C fragment is elevated in patients with COPD compared to healthy controls.

Publication types

  • Validation Study

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Amino Acid Sequence
  • Animals
  • Blood Chemical Analysis / methods*
  • Case-Control Studies
  • Cathepsins / metabolism*
  • Collagen Type III / blood*
  • Collagen Type III / immunology*
  • Collagen Type III / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / analysis*
  • Epitopes / immunology
  • Epitopes / metabolism
  • Female
  • Humans
  • Immunosorbents
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Middle Aged
  • Proteolysis*
  • Pulmonary Disease, Chronic Obstructive / blood
  • Pulmonary Disease, Chronic Obstructive / diagnosis*

Substances

  • Collagen Type III
  • Epitopes
  • Immunosorbents
  • Cathepsins

Grants and funding

This work was supported by the Danish Agency for Science, Technology and Innovation, and the Danish Research Foundation. DGKR, JMBS, MAK, and FGE are employees and MAK is a shareholder of Nordic Bioscience. The funder provided support in the form of salaries for authors DGKR, JMBS, MAK, and FGE, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the 'author contributions' section.