Antibody-selectable filamentous fd phage vectors: affinity purification of target genes

Gene. 1988 Dec 20;73(2):305-18. doi: 10.1016/0378-1119(88)90495-7.

Abstract

Foreign DNA fragments can be inserted into a minor coat protein gene of filamentous phage, creating a fusion protein that is incorporated into the virion; we call these particles "fusion phage". The foreign amino acids are displayed on the surface, allowing fusion phage bearing antigenic determinants from a target gene to be purified in infectious form by affinity to antibody directed against the gene product. Here we introduce fusion-phage vectors that accept foreign DNA inserts with little effect on phage function; and describe affinity purification of virions bearing a target determinant from a 10(8)-fold excess of phage not bearing the determinant, using minute amounts of antibody. These "antibody-selectable" vectors are a promising alternative to conventional expression systems for using antibodies to clone genes, though the ability to isolate rare clones from actual libraries remains to be demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies
  • Base Sequence
  • Chromatography, Affinity
  • Cloning, Molecular / methods
  • Coliphages / drug effects
  • Coliphages / genetics*
  • DNA, Viral / genetics*
  • DNA, Viral / immunology
  • Escherichia coli / genetics
  • Genes, Viral*
  • Genetic Vectors*
  • Molecular Sequence Data
  • Transduction, Genetic
  • Ultraviolet Rays
  • Virion / genetics
  • beta-Galactosidase / genetics

Substances

  • Antibodies
  • DNA, Viral
  • beta-Galactosidase