The affinity-selection of a minibody polypeptide inhibitor of human interleukin-6

Franck Martin, Carlo Toniatti, Anna Laura Salvati, Sara Venturini, Gennaro Ciliberto, Riccardo Cortese, Maurizio Sollazzo

Research output: Contribution to journalArticle

Abstract

A major challenge in basic and applied biological research is the engineering of small proteins with predetermined structures and novel functions. In a limited number of cases, this has been achieved by de novo design. An alternative combinatorial approach is based on the construction of large libraries of random peptides and on methods for the selection of the desired molecules. Here we describe a successful combination of both the rational design and the combinatorial approaches for developing proteins with useful biological functions, in this case the construction of a specific inhibitor of the cytokine human interleukin-6. In previous work, the 'minibody', a 61 residue polypeptide consisting of a β-pleated framework and two hypervariable regions, was designed, synthesized and expressed on f1 phage surface. We report the construction of a repertoire of 50 million minibodies displayed on phage in which the hypervariable regions have been randomized. One polypeptide which binds tightly and specifically to human interleukin-6 was isolated from this collection of minibody mutants. This particular minibody is an effective inhibitor of the cytokine's biological activity. The approach described here could in principle be applied to other molecular targets.

Original languageEnglish
Pages (from-to)5303-5309
Number of pages7
JournalEMBO Journal
Volume13
Issue number22
Publication statusPublished - Nov 15 1994

Keywords

  • Affinity-selection
  • Constrained peptides
  • Human IL-6 inhibitor
  • Phage display
  • Protein engineering

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

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    Martin, F., Toniatti, C., Salvati, A. L., Venturini, S., Ciliberto, G., Cortese, R., & Sollazzo, M. (1994). The affinity-selection of a minibody polypeptide inhibitor of human interleukin-6. EMBO Journal, 13(22), 5303-5309.