Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector

Franco Felici, Luisa Castagnoli, Andrea Musacchio, Roberto Jappelli, Gianni Cesareni

Research output: Contribution to journalArticle

368 Citations (Scopus)

Abstract

Practically any oligopeptide can be exposed on the surface of the bacteriophage capsid by fusion to the major coat protein of filamentous bacteriophages. A phage expressing a particular peptide tag can be selected from a mixture of tens of millions of clones, exposing oligopeptides of random sequence, by affinity purification with a protein ligand. In this respect, pVIII can be used as an alternative and complement to the exposition vectors based on the product of gene III (pIII). We have constructed a phagemid vector that contains gene VIII under the control of the pLac promoter. This vector can be conveniently used to construct libraries of oligopeptides with a random amino acid sequence. An antipeptide monoclonal antibody was used to affinity-purify phagemids exposing oligopeptides which can interact with the monoclonal antibody. DNA sequencing of the amino terminus of gene VIII of the recovered clones predicts the synthesis of hybrid proteins whose amino-terminal amino acid sequence is related to that of the oligopeptide used to raise the antibody. In other words, only oligopeptides that bind a very small portion of the immunoglobulin G surface are affinity-purified by this method, implying that the antigen binding site possesses molecular properties that renders it much stickier than the remainder of the molecule.

Original languageEnglish
Pages (from-to)301-310
Number of pages10
JournalJournal of Molecular Biology
Volume222
Issue number2
DOIs
Publication statusPublished - Nov 20 1991

Fingerprint

Oligopeptides
Ligands
Antibodies
Bacteriophages
Amino Acid Sequence
Inovirus
Clone Cells
Monoclonal Antibodies
Genes
Capsid
Capsid Proteins
DNA Sequence Analysis
Proteins
Immunoglobulin G
Binding Sites
Antigens
Peptides

Keywords

  • filamentous phages
  • IL1β
  • major capsid protein
  • peptide library

ASJC Scopus subject areas

  • Virology

Cite this

Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. / Felici, Franco; Castagnoli, Luisa; Musacchio, Andrea; Jappelli, Roberto; Cesareni, Gianni.

In: Journal of Molecular Biology, Vol. 222, No. 2, 20.11.1991, p. 301-310.

Research output: Contribution to journalArticle

Felici, Franco ; Castagnoli, Luisa ; Musacchio, Andrea ; Jappelli, Roberto ; Cesareni, Gianni. / Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. In: Journal of Molecular Biology. 1991 ; Vol. 222, No. 2. pp. 301-310.
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