Development and application of fully functional epitope-tagged forms of transforming growth factor-γ

Lawrence A. Wolfraim, Gonnie M. Alkemade, Biju Alex, Shellyann Sharpe, W. Tony Parks, John J. Letterio

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Administration of transforming growth factor-β (TGF-β) has been found to be of therapeutic benefit in various mouse disease models and has potential clinical usefulness. However, the ability to track the distribution of exogenously administered, recombinant forms of these proteins has been restricted by cross-reactivity with endogenous TGF-β and related TGF-β isoforms. We describe novel FLAG- and hemagglutinin (HA)-tagged versions of mature TGF-β1 that retain full biological activity as demonstrated by their ability to inhibit the growth of Mv1Lu epithelial cells, and to induce phosphorylation of the TGF-β signaling intermediate, smad 2. Intracellular FLAG- and HA-TGF-β1 can be detected in transfected cells by confocal immunofluorescence microscopy. We also describe sandwich ELISAs designed to specifically detect epitope-tagged TGF-β and demonstrate the utility of these tagged ligands as probes for TGF-β receptor expression by flow cytometry. The design of these fully functional epitope-tagged TGF-β proteins should facilitate studies such as the evaluation of in vivo peptide pharmacodynamics and trafficking of TGF-β ligand-receptor complexes.

Original languageEnglish (US)
Pages (from-to)7-18
Number of pages12
JournalJournal of Immunological Methods
Volume266
Issue number1-2
DOIs
StatePublished - Aug 1 2002

Keywords

  • Biological activity
  • Epitope tag
  • TGF-β

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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