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 potentia...

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Bibliographic Details
Published inNature biotechnology Vol. 16; no. 11; pp. 1068 - 1073
Main Authors Monaci, Paolo, Bartoli, Fabrizia, Nuzzo, Maurizio, Urbanelli, Lorena, Bellintani, Francesca, Prezzi, Caterina, Cortese, Riccardo
Format Journal Article
LanguageEnglish
Published New York, NY Nature 01.11.1998
Subjects
Amino Acid Sequence
Antigens, Viral - genetics
Biological and medical sciences
Biotechnology
Chromosome Mapping
DNA - genetics
DNA Primers - genetics
Fundamental and applied biological sciences. Psychology
Genes, Viral
Health. Pharmaceutical industry
Hepacivirus - genetics
Hepacivirus - immunology
Hepatitis C - immunology
Hepatitis C Antibodies - blood
Hepatitis C Antibodies - metabolism
Humans
Immunological technics applied to diagnosis
In Vitro Techniques
Industrial applications and implications. Economical aspects
Ligands
Molecular Sequence Data
Nucleic Acid Hybridization
Peptide Library
Peptides - genetics
Peptides - immunology
Polymerase Chain Reaction
Virus
Polymerase chain reaction
Molecular hybridization
Peptide library
Specificity
Antibody
Ligand
Diagnosis
Method
Flaviviridae
Hepatitis C virus
Hepacivirus
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Summary: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.
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ISSN:1087-0156
1546-1696
DOI:10.1038/3525