Use of living columns to select specific phage antibodies.
Here we demonstrate that it is possible to confront two recombinant microorganisms in order to select one using the other. We have shown that an epitope derived from p21ras expressed within the outer membrane protein, LamB, can be recognized both by the monoclonal antibody Y13-259, as well as the single chain Fv fragment derived from it. This specificity, which is maintained when the Y13-259 single chain Fv is expressed as a fusion protein with the phage fd gene 3 protein, has allowed us to use the living column of LamB-ras to purify Y13-259 phage from a background of non-binding phage, even at dilutions as high as 10 phage in 10(10) irrelevant phage.
Bradbury
A
A
SISSA, Biophysics Sector, Trieste, Italy.
Persic
L
L
Werge
T
T
Cattaneo
A
A
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D005656:Fungal Proteins / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000276:immunology*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005656:Fungal Proteins; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras)