Merck

Protocol for design, construction, and selection of genome phage (gPhage) display libraries.

STAR protocols (2021-11-23)
Luis Antonio Rodriguez Carnero, André Azevedo Reis Teixeira, Fenny Hui Fen Tang, Andréia Kuramoto, Maria Júlia Manso Alves, Walter Colli, João Carlos Setubal, Edécio Cunha-Neto, Renata Pasqualini, Wadih Arap, Ricardo José Giordano
RESUMEN

This protocol describes the genomic phage (gPhage) display platform, a large-scale antigen and epitope mapping technique. We constructed a gPhage display peptide library of a eukaryotic organism, Trypanosoma cruzi (causative agent of Chagas disease), to map the antibody response landscape against the parasite. Here, we used an organism with a relatively large but intronless genome, although future applications could include other prevalent or (re)emerging infectious organisms carrying genomes with a limited number of introns. For complete details on the use and execution of this protocol, please refer to Teixeira et al. (2021).

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Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)
Sigma-Aldrich
Glycerol, for molecular biology, ≥99.0%
Sigma-Aldrich
SIGMAFAST OPD, tablet