Development of an enzyme-linked immunosorbent assay for measurement of rat pulmonary surfactant protein D using monoclonal antibodies

Exp Lung Res. 2010 Oct;36(8):463-8. doi: 10.3109/01902141003746371.

Abstract

Surfactant protein D (SP-D) has been used as a biomarker of lung inflammation. In rat, several types of enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies have been reported. The purpose of this study was the development of a sensitive ELISA for rat SP-D using monoclonal antibodies. The authors developed a sandwich ELISA using monoclonal antibodies that were obtained by immunizing with purified rat SP-D. The ELISA was evaluated by performance tests. Furthermore, concentrations of serum SP-D were measured in normal control and bleomycin-treated rats. The working range of ELISA was between 0.47 and 30 ng/mL. Different concentrations of added SP-D were recovered, between 94.1% and 102.8%. Serum SP-D levels of bleomycin-treated rats were significantly higher than those of normal rats. In conclusion, this newly developed ELISA for rat SP-D using monoclonal antibodies is applicable for research on the mechanism and therapy of lung injury.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology*
  • Bleomycin / toxicity
  • Blotting, Western
  • Bronchoalveolar Lavage Fluid / chemistry
  • Disease Models, Animal
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Lung / drug effects
  • Lung / metabolism*
  • Male
  • Pulmonary Surfactant-Associated Protein D / analysis*
  • Pulmonary Surfactant-Associated Protein D / blood
  • Pulmonary Surfactant-Associated Protein D / immunology
  • Rats
  • Rats, Inbred F344
  • Rats, Wistar

Substances

  • Antibodies, Monoclonal
  • Pulmonary Surfactant-Associated Protein D
  • Bleomycin