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

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

doi:10.1016/j.xpro.2021.100936

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

STAR Protoc. 2021 Nov 8;2(4):100936. doi: 10.1016/j.xpro.2021.100936. eCollection 2021 Dec 17.

Authors

Luis Antonio Rodriguez Carnero¹, André Azevedo Reis Teixeira¹, Fenny Hui Fen Tang^{1

2}, Andréia Kuramoto³, Maria Júlia Manso Alves¹, Walter Colli¹, João Carlos Setubal¹, Edécio Cunha-Neto^{3

4

5}, Renata Pasqualini^{2

6}, Wadih Arap^{2

7}, Ricardo José Giordano^{1

5}

Affiliations

¹ Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP 05508, Brazil.
² Rutgers Cancer Institute of New Jersey, Newark, NJ 07101, USA.
³ Heart Institute (InCor), University of São Paulo School of Medicine, São Paulo, SP 05403, Brazil.
⁴ Division of Clinical Immunology and Allergy, University of São Paulo School of Medicine, São Paulo, SP 01246, Brazil.
⁵ Institute for Investigation in Immunology, University of São Paulo School of Medicine, São Paulo, SP 01246, Brazil.
⁶ Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ 07103, USA.
⁷ Division of Hematology/Oncology, Department of Medicine, Rutgers New Jersey Medical School, Newark, NJ 07103, USA.

Abstract

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).

Keywords: Antibody; Genomics; High Throughput Screening; Immunology; Microbiology; Model Organisms; Molecular Biology; Molecular/Chemical Probes; Sequence analysis; Sequencing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Protozoan / chemistry
Antibodies, Protozoan / metabolism
Cell Surface Display Techniques / methods*
Genome, Protozoan / genetics
Genomic Library*
Trypanosoma cruzi / genetics

Substances

Antibodies, Protozoan