Antibody-selectable filamentous fd phage vectors: affinity purification of target genes.
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.
Parmley
S F
SF
Division of Biological Sciences, University of Missouri, Columbia 65211.
Smith
G P
GP
D000906:Antibodies; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods; D003090:Coliphages / Q000187:drug effects / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D014161:Transduction, Genetic; D014466:Ultraviolet Rays; D014771:Virion / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D000906:Antibodies; D004279:DNA, Viral; D001616:beta-Galactosidase