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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.

SUBMITTER: Monaci P

PROVIDER: S-EPMC2204054 | biostudies-literature | 2008 Jan

REPOSITORIES: biostudies-literature

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Differential screening of phage-ab libraries by oligonucleotide microarray technology.

Monaci Paolo P Luzzago Alessandra A Santini Claudia C De Pra Alessandra A Arcuri Mirko M Magistri Francesca F Bellini Alessandro A Ansuini Helenia H Ambrosio Maria M Ammendola Virginia V Bigotti Maria Giulia MG Cirillo Agostino A Nuzzo Maurizio M Nasti Annamaria Assunta AA Neuner Philippe P Orsatti Laura L Pezzanera Monica M Sbardellati Andrea A Silvestre Giuseppe G Uva Paolo P Viti Valentina V Barbato Gaetano G Colloca Stefano S Demartis Anna A De Rinaldis Emanuele E Giampaoli Saverio S Lahm Armin A Palombo Fabio F Talamo Fabio F Vitelli Alessandra A Nicosia Alfredo A Cortese Riccardo R

PloS one 20080130 1

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 ex ...[more]

PMID: 18231595

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Differential screening of phage-ab libraries by oligonucleotide microarray technology.

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Differential screening of phage-ab libraries by oligonucleotide microarray technology.

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