1. Biopanning and rapid analysis of selective interactive ligands
- Author
-
Ricardo J. Giordano, Renata Pasqualini, Marina Cardó-Vila, Johanna Lahdenranta, and Wadih Arap
- Subjects
Vascular Endothelial Growth Factor A ,Recombinant Fusion Proteins ,Amino Acid Motifs ,Molecular Sequence Data ,Peptide ,Nerve Tissue Proteins ,Biopanning ,Endothelial Growth Factors ,Biology ,Ligands ,General Biochemistry, Genetics and Molecular Biology ,chemistry.chemical_compound ,Peptide Library ,Neuropilin 1 ,Humans ,Receptors, Growth Factor ,Amino Acid Sequence ,Peptide library ,Peptide sequence ,Cells, Cultured ,chemistry.chemical_classification ,Differential centrifugation ,Lymphokines ,Vascular Endothelial Growth Factors ,Cell Membrane ,Receptor Protein-Tyrosine Kinases ,General Medicine ,Molecular biology ,Neuropilin-1 ,Vascular endothelial growth factor ,Vascular endothelial growth factor A ,Receptors, Vascular Endothelial Growth Factor ,Biochemistry ,chemistry ,Endothelium, Vascular ,Peptides - 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.
- Published
- 2001