DNA-based selection and screening of peptide ligands

Nat Biotechnol. 1998 Nov;16(11):1068-73. doi: 10.1038/3525.

Abstract

Phage display selection strategies rely on the physical link between the displayed heterologous protein ligand and the DNA encoding it. Thus, genes expressing a ligand with a specific binding affinity can be selected rapidly. To improve the specificity and sensitivity of this technology for potential use in identifying ligands to a specific antibody present in a complex mixture, we incorporated a DNA selection step along with the phage display technology. Ligands for hepatitis C virus (HCV) antibodies present in serum were identified by panning a phage-displayed random peptide library against pools of serum HCV antibodies. An additional DNA hybridization screening step using single-stranded DNA isolated from one of the pools increased the specificity and sensitivity, resulting in the selection of an HCV antibody ligand with diagnostic potential.

MeSH terms

  • Amino Acid Sequence
  • Antigens, Viral / genetics
  • Biotechnology
  • Chromosome Mapping
  • DNA / genetics*
  • DNA Primers / genetics
  • Genes, Viral
  • Hepacivirus / genetics
  • Hepacivirus / immunology
  • Hepatitis C / immunology
  • Hepatitis C Antibodies / blood
  • Hepatitis C Antibodies / metabolism
  • Humans
  • In Vitro Techniques
  • Ligands
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Peptide Library
  • Peptides / genetics*
  • Peptides / immunology*
  • Polymerase Chain Reaction

Substances

  • Antigens, Viral
  • DNA Primers
  • Hepatitis C Antibodies
  • Ligands
  • Peptide Library
  • Peptides
  • DNA