Abstract
Size and variability often represent an obstacle in generating an effective antibody gene library for the detection of an abundant repertoire of antigens. Therefore, optimizing the construction of a large library is essential for the selection of high-affinity reactive fragments. Here, we report a highly efficient method for the construction of a human naïve antibody gene library for the selection of antibodies as single-chain variable fragments. This protocol is based on many different sets of oligonucleotide primers and multistep electroporation and ligation reactions.
This advanced method can be adopted by any molecular biology laboratory to generate a naïve library for use in isolating single-chain fragment variables against specific targets.
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Acknowledgment
This work was supported by the Italian Association for Cancer Research (IG2016_18451 to KS) and by a grant from the Regione Emilia Romagna Project: POR-FESR2007-2013 (PR WEB IOR 1654/2010).
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Pasello, M., Mallano, A., Flego, M., Zamboni, S., Giudice, A.M., Scotlandi, K. (2018). Construction of Human Naïve Antibody Gene Libraries. In: Nevoltris, D., Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 1827. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8648-4_4
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DOI: https://doi.org/10.1007/978-1-4939-8648-4_4
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