Preparation of phage antibodies to the ED-A domain of human fibronectin

Laura Borsi, Patrizia Castellani, Giorgio Allemanni, Dario Neri, Luciano Zardi

Research output: Contribution to journalArticle

Abstract

The fibronectin (FN) isoform containing the alternative spliced ED-A domain is much more expressed in fetal, tumoral, and regenerating tissues than in normal adult tissues. The ED-A containing FN is up-regulated by numerous cytokines, such as TGF-β, and, although in normal adult liver the ED-A domain is undetectable, in regenerating rat liver the expression of ED- A is increased and mediates the conversion of fat storing cells to myofibroblasts. Here we describe the selection from a phage display library and the characterization of human antibody fragments directed against the EDA sequence of FN. As they can be easily radiolabeled with 32P, these antibodies are very highly sensitive reagents for the determination of ED-A levels in tissues and biological fluids; in fact, use of these scFv induced a more than 10-fold increase in sensitivity with respect to the murine monoclonal IST-9. The possibility of preparing a range of human engineered antibodies should facilitate the development of antibody reagents with suitable pharmacokinetics, valency, functional affinity, and effector functions and that could be useful for clinical purposes.

Original languageEnglish
Pages (from-to)244-251
Number of pages8
JournalExperimental Cell Research
Volume240
Issue number2
DOIs
Publication statusPublished - May 1 1998

ASJC Scopus subject areas

  • Cell Biology

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