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Screening Phage-Display Antibody Libraries Using Protein Arrays

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Phage Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

Phage-display technology constitutes a powerful tool for the generation of specific antibodies against a predefined antigen. The main advantages of phage-display technology in comparison to conventional hybridoma-based techniques are: (1) rapid generation time and (2) antibody selection against an unlimited number of molecules (biological or not). However, the main bottleneck with phage-display technology is the validation strategies employed to confirm the greatest number of antibody fragments. The development of new high-throughput (HT) techniques has helped overcome this great limitation. Here, we describe a new method based on an array technology that allows the deposition of hundreds to thousands of phages by micro-contact on a unique nitrocellulose surface. This setup comes in combination with bioinformatic approaches that enables simultaneous affinity screening in a HT format of antibody-displaying phages.

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Acknowledgments

We gratefully acknowledge financial support from the Carlos III Health Institute of Spain (FIS PI14/01538), Fondos FEDER (EU), Junta Castilla-Leon (BIO/SA07/15), and Fundación Solórzano FS/23-2015. The proteomics Unit belongs to ProteoRed-ISCIII, PRB2-ISCII, supported by grant PT13/001. P.D. is supported by a JCYL-EDU/346/2013 Ph.D. scholarship.

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Authors and Affiliations

  1. ImmunoStep SL. Edificio Centro de Investigación del Cáncer. Avda. Coimbra s/n, 37007, Salamanca, Spain

    Ricardo Jara-Acevedo

  2. Department of Medicine and General Cytometry Service-Nucleus, Cancer Research Center (CSIC/USAL/IBSAL), Francisco Vitoria 6-16, 37007, Salamanca, Spain

    Paula Díez, María González-González, Rafael Góngora, Alberto Orfao & Manuel Fuentes

  3. Proteomics Unit, Cancer Research Center (CSIC/USAL/IBSAL), 37007, Salamanca, Spain

    Paula Díez, María González-González, Rosa María Dégano, Nieves Ibarrola & Manuel Fuentes

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  1. Ricardo Jara-Acevedo
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  2. Paula Díez
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  3. María González-González
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  4. Rosa María Dégano
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  5. Nieves Ibarrola
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  6. Rafael Góngora
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  7. Alberto Orfao
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  8. Manuel Fuentes
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Correspondence to Manuel Fuentes .

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Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Jara-Acevedo, R. et al. (2018). Screening Phage-Display Antibody Libraries Using Protein Arrays. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

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