Monoclonal antibodies to the collagen binding domain of human plasma fibronectin

Guido Tarone, Maria Rosa Amedeo, Maria Flavia Di Renzo, Paolo Comoglio

Research output: Contribution to journalArticlepeer-review

Abstract

Five independent hybrids producing monoclonal antibodies to human plasma fibronectin have been obtained by fusing P3/X63-Ag8 myeloma cells with immune mouse splenocytes. The specificity of these monoclonal antibodies (MABs) for fibronectin was demonstrated by three independent tests: Binding to the purified soluble molecule, immunofluorescence staining of insoluble extracellular matrices produced by endothelial cells in vitro, immunostaining of fibronectin tryptic peptides after separation on SDS-PAGE and transfer to nitrocellulose sheets. Two antibodies (MAB 29 and 52) recognized selectively human fibronectin while the others (MAB 5, 30 and 59) reacted also with plasma fibronectin from calf, hamster and chicken. Four distinct epitopes were recognized by the MABs studied. MAB 5, 30, 52 and 59 reacted with distinct antigenic sites, while MAB 29 and 52 bind to the same site. Antigenic fragments were identified by immunostaining of fibronectin tryptic peptides. MAB 5 reacted with a collagen binding fragment with a molecular weight of 120 K. In addition, each of the MAB 29, 30, 52 and 59 reacted with peptides with a molecular weight of 40 K that bind to gelatin. Since these antibodies do not inhibit fibronectin-collagen interaction, it is concluded that their corresponding epitopes are clustered in a region close, but not coincident, to the collagen binding site of fibronectin.

Original languageEnglish
Pages (from-to)225-236
Number of pages12
JournalPathobiology
Volume52
Issue number4
DOIs
Publication statusPublished - 1984

Keywords

  • Collagen
  • Fibronectin
  • Monoclonal antibodies

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pathology and Forensic Medicine

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