Biopanning and rapid analysis of selective interactive ligands

R. J. Giordano, M. Cardó-Vila, J. Lahdenranta, Renata Pasqualini, Wadih Arap

Research output: Contribution to journalArticle

212 Scopus citations

Abstract

Here we introduce a new approach for the screening, selection and sorting of cell-surface-binding peptides from phage libraries. Biopanning and rapid analysis of selective interactive ligands (termed BRASIL) is based on differential centrifugation in which a cell suspension incubated with phage in an aqueous upper phase is centrifuged through a non-miscible organic lower phase. This single-step organic phase separation is faster, more sensitive and more specific than current methods that rely on washing steps or limiting dilution. As a proof-of-principle, we screened human endothelial cells stimulated with vascular endothelial growth factor (VEGF) and constructed a peptide-based ligand-receptor map of the VEGF family. Next, we validated the motif PQPRPL as a novel chimeric ligand mimic that binds specifically to VEGF receptor-1 and to neuropilin-1. BRASIL may prove itself a superior method for probing target cell surfaces with a broad range of potential applications.

Original languageEnglish (US)
Pages (from-to)1249-1253
Number of pages5
JournalNature Medicine
Volume7
Issue number11
DOIs
StatePublished - Nov 26 2001
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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