title: Isolation of human prostate cancer cell reactive antibodies using phage display technology.
abstract: Here we describe a phage display strategy for the selection of rabbit monoclonal antibodies that recognize cell surface tumor-associated antigens expressed in prostate cancer. Two immune rabbit/human chimeric Fab libraries were displayed on phage and used to search for tumor-associated antigens by panning on DU145 human prostate cancer cells. For this, we developed a novel whole-cell panning protocol with two negative selection steps designed to remove antibodies reacting with common antigens. After three rounds of subtractive panning, a majority of clones bound to DU145 cells as detected by flow cytometry. Among these, we identified several clones that bound selectively to DU145 cells but not to primary human prostate epithelial cell line PrEC. In summary, our work demonstrates the potential of immune rabbit antibody libraries for target discovery in general and the identification of cell surface tumor-associated antigens in particular.
authors.affiliation: The Skaggs Institute for Chemical Biology and the Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Popkov
authors.forename: Christoph
authors.initials: C
authors.lastname: Rader
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D016172:DNA Fingerprinting; D005434:Flow Cytometry; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D008297:Male; D019151:Peptide Library; D011471:Prostatic Neoplasms / Q000235:genetics / Q000276:immunology*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011506:Proteins