Abstract
The advent of the serological identification of antigens by procedures such as cDNA cloning and recombinant protein expression has allowed the direct molecular definition of immunogenic proteins. The phage-display technology provides several advantages over conventional immunoscreening procedures based on plasmid or lambda-phage cDNA libraries. So far, attempts to display open reading frames, such as those encoded by cDNA fragments, on filamentous phages have not been very successful. We managed to develop a strategy based on “folding reporters” which allows filtering out open reading frames from DNA and displaying them on filamentous phages in such a way that they are amenable to subsequent selection or screening.
Once the cDNA library of interest is created, phage-display technology is used for selection of novel putative antigens; these are then validated by printing isolated protein on microarray and screening with patients’ sera.
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Acknowledgements
This work was supported in part by Fondazione Cariplo, Compagnia Sanpaolo, NIH RFA-DK-06-002, and Regione Piemonte Ricerca sanitaria Finalizzata 2006 and 2007.
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Di Niro, R., D’Angelo, S., Secco, P., Marzari, R., Santoro, C., Sblattero, D. (2009). Profiling the Autoantibody Repertoire by Screening Phage-Displayed Human cDNA Libraries. In: Cretich, M., Chiari, M. (eds) Peptide Microarrays. Methods in Molecular Biology™, vol 570. Humana Press. https://doi.org/10.1007/978-1-60327-394-7_20
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DOI: https://doi.org/10.1007/978-1-60327-394-7_20
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