epitope description:SDLWK

antigen name:Cellular tumor antigen p53

host organism:Mus musculus

antibody name:p53-11

instance of dataset

available

registration not required

Literature

J Immunol Methods

1997

F Fack

B Hü

gle-Dö

rr

D Song

I Queitsch

G Petersen

E K Bautz

phage display

Epitope mapping by phage display: random versus gene-fragment libraries.

Epitope found to be recognized by an anti-p53 mAb by panning of phage display libraries, either of gene fragments or of random hexapeptides, followed by overlapping peptides scan.

Fragments of the p53 protein expressed onto a phage display library at the N-terminal end of pIII protein were selected with an anti-p53 mAb thought to recognize a linear epitope in that it interacts with the protein on immunoblots. After one round of panning 65% of clones were positive by ELISA . Sequencing of the positive clones reavealed the presence of 7 clones, which overlap in a 12-aa region that contains the epitope. Only one of those clones sequence is shown here. Comparative use of a random 6-mer peptide phage display library allowed for the selection of 85% of positive binders by ELISA after 4 rounds. Sequencing of the positive clones revealed 1 unique insert (YSDLGK) which includes 4 residues of the 12-mer found by gene-fragments panning. Panning of a 15-mer random library allowed for the selection of 100% of positive binders after 4 rounds. Sequencing of the positive clones revealed 1 unique insert (TDAFSDLGRMLANPG) which includes 5 residues of the 12-mer found by gene-fragments panning.

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