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High-throughgput phage-display screening in array format

AutorDíez, Paula; Jara-Acevedo, Ricardo; González González, María; Casado-Vela, Juan; Dasilva, Noelia; Lecrevisse, Q.; Bartolomé, Raquel; Orfao, Alberto; Fuentes, Manuel
Fecha de publicación2015
CitaciónEnzyme and Microbial Technology 79-80: 34-41 (2015)
ResumenEmerging technologies for the design and generation of human antibodies require improved approaches enabling their screening, characterization and validation. Currently, strategies based on ELISA or western blot are used to that aim. However, the ever increasing number of novel antibodies generated would benefit from the development of new high-throughput (HT) platforms facilitating rapid antibody identification and characterization. Herein, we describe a protein chip bearing recombinant phage particles and based on a large phage antibody library. In this paper we have set forth a novel implementation which provides a powerful and simple methodology enabling the identification of single-chain variable fragments (scFv). As a proof-of-principle of this method, we tested it with recombinant antigen (human recombinant interleukin 8). Additionally, we developed a novel bioinformatics tool that serves to compare this novel strategy with traditional methods. The method described here, together with associated informatics tools, is robust, relatively fast and represents a step-forward in protocols including phage library screenings.
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Identificadoresdoi: 10.1016/j.enzmictec.2015.06.016
issn: 0141-0229
e-issn: 1879-0909
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