Biopanning and rapid analysis of selective interactive ligands

Nat Med. 2001 Nov;7(11):1249-53. doi: 10.1038/nm1101-1249.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Endothelial Growth Factors / metabolism
  • Endothelial Growth Factors / pharmacology
  • Endothelium, Vascular / metabolism
  • Humans
  • Ligands
  • Lymphokines / metabolism
  • Lymphokines / pharmacology
  • Molecular Sequence Data
  • Nerve Tissue Proteins / metabolism
  • Neuropilin-1
  • Peptide Library*
  • Peptides / chemistry
  • Peptides / metabolism
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptors, Growth Factor / metabolism
  • Receptors, Vascular Endothelial Growth Factor
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors

Substances

  • Endothelial Growth Factors
  • Ligands
  • Lymphokines
  • Nerve Tissue Proteins
  • Peptide Library
  • Peptides
  • Receptors, Growth Factor
  • Recombinant Fusion Proteins
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • Neuropilin-1
  • Receptor Protein-Tyrosine Kinases
  • Receptors, Vascular Endothelial Growth Factor