Characterization of prostate-specific antigen binding peptides selected by phage display technology.
Prostate-specific antigen (PSA) is an important marker for the diagnosis and management of prostate cancer. Free PSA has been shown to be more extensively cleaved in sera from benign prostatic hyperplasia patients than in sera from prostate cancer patients. Moreover, the presence of enzymatically activatable PSA was characterized previously in sera from patients with prostate cancer by the use of the specific anti-free PSA monoclonal antibody (mAb) 5D3D11. As an attempt to obtain ligands for the specific recognition of different PSA forms including active PSA, phage-displayed linear and cyclic peptide libraries were screened with PSA coated directly into microplate wells or presented by two different anti-total PSA mAbs. Four different phage clones were selected for their ability to recognize PSA and the inserted peptides were produced as synthetic peptides. These peptides were found to capture and to detect specifically free PSA, even in complex biological media such as sera or tumour cell culture supernatants. Alanine scanning of peptide sequences showed the involvement of aromatic and hydrophobic residues in the interaction of the peptides with PSA whereas Spotscan analysis of overlapping peptides covering the PSA sequence identified a peptide binding to the kallikrein loop at residues 82-87, suggesting that the peptides could recognize a non-clipped form of PSA. Moreover, the PSA-specific peptides enhance the enzymatic activity of PSA immobilized into microplate wells whereas the capture of PSA by the peptides inhibited totally its enzymatic activity while the peptide binding to PSA had no effect in solution. These PSA-specific peptides could be potential tools for the recognition of PSA forms more specifically associated to prostate cancer.
Ferrieu-Weisbuch
Catherine
C
Unité Mixte de Recherche UMR 2714 CNRS-bioMérieux, IFR 128 BioSciences Lyon-Gerland, 21 Avenue Tony Garnier, 69365 LYON Cedex 07, France.
Michel
Sandrine
S
Collomb-Clerc
Emilie
E
Pothion
Catherine
C
Deléage
Gilbert
G
Jolivet-Reynaud
Colette
C
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017430:Prostate-Specific Antigen / Q000032:analysis* / Q000097:blood / Q000276:immunology; D011485:Protein Binding
D000911:Antibodies, Monoclonal; D019151:Peptide Library; D010455:Peptides; D017430:Prostate-Specific Antigen