title: Differential screening of phage-ab libraries by oligonucleotide microarray technology.
abstract: 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.
authors.affiliation: Biotechnology Department, Istituto di Ricerca di Biologia Molecolare P. Angeletti, Pomezia, Rome, Italy. paolo_monaci@merck.com
authors.forename: Paolo
authors.initials: P
authors.lastname: Monaci
authors.forename: Alessandra
authors.initials: A
authors.lastname: Luzzago
authors.forename: Claudia
authors.initials: C
authors.lastname: Santini
authors.forename: Alessandra
authors.initials: A
authors.lastname: De Pra
authors.forename: Mirko
authors.initials: M
authors.lastname: Arcuri
authors.forename: Francesca
authors.initials: F
authors.lastname: Magistri
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bellini
authors.forename: Helenia
authors.initials: H
authors.lastname: Ansuini
authors.forename: Maria
authors.initials: M
authors.lastname: Ambrosio
authors.forename: Virginia
authors.initials: V
authors.lastname: Ammendola
authors.forename: Maria Giulia
authors.initials: MG
authors.lastname: Bigotti
authors.forename: Agostino
authors.initials: A
authors.lastname: Cirillo
authors.forename: Maurizio
authors.initials: M
authors.lastname: Nuzzo
authors.forename: Annamaria Assunta
authors.initials: AA
authors.lastname: Nasti
authors.forename: Philippe
authors.initials: P
authors.lastname: Neuner
authors.forename: Laura
authors.initials: L
authors.lastname: Orsatti
authors.forename: Monica
authors.initials: M
authors.lastname: Pezzanera
authors.forename: Andrea
authors.initials: A
authors.lastname: Sbardellati
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Silvestre
authors.forename: Paolo
authors.initials: P
authors.lastname: Uva
authors.forename: Valentina
authors.initials: V
authors.lastname: Viti
authors.forename: Gaetano
authors.initials: G
authors.lastname: Barbato
authors.forename: Stefano
authors.initials: S
authors.lastname: Colloca
authors.forename: Anna
authors.initials: A
authors.lastname: Demartis
authors.forename: Emanuele
authors.initials: E
authors.lastname: De Rinaldis
authors.forename: Saverio
authors.initials: S
authors.lastname: Giampaoli
authors.forename: Armin
authors.initials: A
authors.lastname: Lahm
authors.forename: Fabio
authors.initials: F
authors.lastname: Palombo
authors.forename: Fabio
authors.initials: F
authors.lastname: Talamo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Vitelli
authors.forename: Alfredo
authors.initials: A
authors.lastname: Nicosia
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cortese
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000378:metabolism / Q000493:pharmacokinetics; D001435:Bacteriophages / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D020411:Oligonucleotide Array Sequence Analysis; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008565:Membrane Proteins