Single-chain variable fragments selected on the 57-76 p21Ras neutralising epitope from phage antibody libraries recognise the parental protein.
Phage antibodies have been widely prospected as an alternative to the use of monoclonal antibodies prepared by traditional means. Many monoclonal antibodies prepared against peptides are able to recognise the native proteins from which they were derived. Here we show that the same is also true for phage antibodies. We have selected a number of single-chain variable fragments (scFv) from a large phage scFv library against a peptide from the switch region II of p21Ras. This peptide is known to reside in a mobile area of the native protein and is the epitope of a well characterised monoclonal antibody. Selected scFvs were able to recognise native p21Ras in both ELISA and Western blots, indicating that peptides are also likely to be very useful in selecting from phage antibody libraries.
Persic
L
L
Laboratory of Molecular Neurobiology, International School of Advanced Studies (SISSA), Trieste, Italy.
Horn
I R
IR
Rybak
S
S
Cattaneo
A
A
Hoogenboom
H R
HR
Bradbury
A
A
D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000235:genetics; D015153:Blotting, Western; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016283:Proto-Oncogene Proteins p21(ras) / Q000737:chemistry / Q000276:immunology*