DNA-based selection and screening of peptide ligands.
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.
Bartoli
F
F
Istituto di Ricerche di Biologia Molecolare P. Angeletti, Pomezia (Roma), Italy.
Nuzzo
M
M
Urbanelli
L
L
Bellintani
F
F
Prezzi
C
C
Cortese
R
R
Monaci
P
P
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000235:genetics; D001709:Biotechnology; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005814:Genes, Viral; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology; D006526:Hepatitis C / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D019151:Peptide Library; D010455:Peptides / Q000235:genetics* / Q000276:immunology*; D016133:Polymerase Chain Reaction
D000956:Antigens, Viral; D017931:DNA Primers; D018937:Hepatitis C Antibodies; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D004247:DNA