A competitive enzyme-linked immunosorbent assay for quantification of tetrastatin in body fluids and tumor extracts

Anal Biochem. 2015 Aug 1:482:16-21. doi: 10.1016/j.ab.2015.04.023. Epub 2015 Apr 29.

Abstract

Basement membrane collagens or derived fragments are measured in biological fluids such as blood and urine of patients and appear to be useful for diagnosis, prognostication, or treatment monitoring as proposed for endostatin, a fragment of collagen XVIII, or tumstatin, a fragment of collagen IV. Tetrastatin, the NC1 alpha 4 collagen IV domain, was previously reported to inhibit tumor growth and angiogenesis. The aim of this study was to develop and validate a method to measure tetrastatin concentrations in human fluids. We developed a competitive enzyme-linked immunosorbent assay (ELISA). It allowed measuring tetrastatin levels in human serum, bronchial aspiration and bronchoalveolar lavage fluids, and lung tissue extracts. The tetrastatin level was significantly higher in tumor tissues than in healthy lung tissues. Tetrastatin competitive ELISA could be useful to quantify tetrastatin in tissues and biological fluids for the diagnosis or prognostication of diseases in which basement membrane metabolism may be altered, especially tumor progression.

Keywords: Collagen IV; Competitive ELISA; Pathologies; Tetrastatin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Bronchoalveolar Lavage Fluid / chemistry
  • Child
  • Collagen Type IV / analysis*
  • Collagen Type IV / blood*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Lung / chemistry
  • Lung / pathology*
  • Lung Neoplasms / blood*
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / pathology
  • Male
  • Middle Aged
  • Protein Structure, Tertiary
  • Young Adult

Substances

  • COL4A4 protein, human
  • Collagen Type IV