Differential screening of phage-ab libraries by oligonucleotide microarray technology

• Published in: PloS one Vol. 3; no. 1; p. e1508
• Main Authors: Monaci, Paolo, Luzzago, Alessandra, Santini, Claudia, De Pra, Alessandra, Arcuri, Mirko, Magistri, Francesca, Bellini, Alessandro, Ansuini, Helenia, Ambrosio, Maria, Ammendola, Virginia, Bigotti, Maria Giulia, Cirillo, Agostino, Nuzzo, Maurizio, Nasti, Annamaria Assunta, Neuner, Philippe, Orsatti, Laura, Pezzanera, Monica, Sbardellati, Andrea, Silvestre, Giuseppe, Uva, Paolo, Viti, Valentina, Barbato, Gaetano, Colloca, Stefano, Demartis, Anna, De Rinaldis, Emanuele, Giampaoli, Saverio, Lahm, Armin, Palombo, Fabio, Talamo, Fabio, Vitelli, Alessandra, Nicosia, Alfredo, Cortese, Riccardo
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 30.01.2008; Public Library of Science (PLoS)
• Subjects: Animals; Antibodies; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - metabolism; Antibodies, Monoclonal - pharmacokinetics; Antigens; Apoptosis; B cells; Bacteriophages - genetics; Binding; Biological properties; Biological samples; Biotechnology; Cancer; Cell proliferation; Cell surface; Cloning; Computational Biology; Deoxyribonucleic acid; DNA; DNA microarrays; Enzyme-Linked Immunosorbent Assay; Genomes; Hybridization; Identification methods; Immunoglobulin G - metabolism; Immunoglobulins; Immunology; Libraries; Library collections; Medical screening; Membrane Proteins - metabolism; Mice; Mice, Inbred BALB C; Molecular Biology; Nucleotide sequence; Oligonucleotide Array Sequence Analysis; Oligonucleotides; Oncology/Gastrointestinal Cancers; Phages; Protein expression; Proteins; Receptors; Surface Plasmon Resonance; Technology; Tissues; Tumor cells; Tumors; Italy
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0001508

A novel and efficient tagArray technology was developed that allows rapid identification of antibodies which bind to receptors with a specific expression profile, in the absence of biological information. This method is based on the cloning of a specific, short nucleotide sequence (tag) in the phagemid coding for each phage-displayed antibody fragment (phage-Ab) present in a library. In order to set up and validate the method we identified about 10,000 different phage-Abs binding to receptors expressed in their native form on the cell surface (10 k Membranome collection) and tagged each individual phage-Ab. The frequency of each phage-Ab in a given population can at this point be inferred by measuring the frequency of its associated tag sequence through standard DNA hybridization methods. Using tiny amounts of biological samples we identified phage-Abs binding to receptors preferentially expressed on primary tumor cells rather than on cells obtained from matched normal tissues. These antibodies inhibited cell proliferation in vitro and tumor development in vivo, thus representing therapeutic lead candidates.