Abstract
Phage display is a powerful tool to produce recombinant antibodies against a given antigen without animal immunization. This technology employs libraries of recombinant bacteriophages that display billions of different functional antibody fragments on their surface. They are selected by panning in vitro against the target antigen in search for specific binders. In this chapter, we describe the selection of single chain variable fragment (scFv) antibodies to be used for detection of allergenic proteins from nuts in food products. The artificial libraries TomLinson I+J (MRC Laboratory of Molecular Biology and MRC Centre for Protein Engineering) were employed that resulted in successful phage-ELISA systems for detection of almond and walnut proteins in commercial food products.
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References
Curzer HJ, Perry G, Wallace MC, Perry D (2016) The three rs of animal research: what they mean for the institutional animal care and use committee and why. Sci Eng Ethics 22:549–565. doi:10.1007/s11948-015-9659-8
Qi H, Lu H, Qiu H-J et al (2012) Phagemid vectors for phage display: properties, characteristics and construction. J Mol Biol 417:129–143. doi:10.1016/j.jmb.2012.01.038
McCafferty J, Griffiths AD, Winter G, Chiswell DJ (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554. doi:10.1038/348552a0
Georgieva Y, Konthur Z (2011) Design and screening of M13 phage display cDNA libraries. Molecules 16:1667–1681. doi:10.3390/molecules16021667
de la Cruz S, Cubillos-Zapata C, López-Calleja IM et al (2015) Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products. Food Control 54:322–330. doi:10.1016/j.foodcont.2015.02.011
Bird RE, Hardman KD, Jacobson JW et al (1988) Single-chain antigen-binding proteins. Science 242:423–426
Kristensen P, Winter G (1998) Proteolytic selection for protein folding using filamentous bacteriophages. Fold Des 3:321–328. doi:10.1016/S1359-0278(98)00044-3
Lee CMY, Iorno N, Sierro F, Christ D (2007) Selection of human antibody fragments by phage display. Nat Protoc 2:3001–3008. doi:10.1038/nprot.2007.448
Weber M, Bujak E, Putelli A et al (2014) A highly functional synthetic phage display library containing over 40 billion human antibody clones. PLoS One 9:e100000. doi:10.1371/journal.pone.0100000
de la Cruz S, López-Calleja IM, Alcocer M et al (2013) Selection of recombinant antibodies by phage display technology and application for detection of allergenic Brazil nut (Bertholletia excelsa) in processed foods. J Agric Food Chem 61:10310–10319. doi:10.1021/jf403347t
Rouet R, Lowe D, Dudgeon K et al (2012) Expression of high-affinity human antibody fragments in bacteria. Nat Protoc 7:364–373. doi:10.1038/nprot.2011.448
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Madrid, R., de la Cruz, S., García, A., Martín, R., González, I., García, T. (2017). Detection of Food Allergens by Phage-Displayed Produced Antibodies. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_9
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DOI: https://doi.org/10.1007/978-1-4939-6925-8_9
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