title: A high-performance thioredoxin-based scaffold for peptide immunogen construction: proof-of-concept testing with a human papillomavirus epitope.
abstract: Escherichia coli thioredoxin has been previously exploited as a scaffold for the presentation/stabilization of peptide aptamers as well as to confer immunogenicity to peptide epitopes. Here we focused on other key features of thioredoxin that are of general interest for the production of safer and more effective peptide immunogens, such as a high thermal stability, lack of cross-reactivity and a low-cost of production. We identified thioredoxin from the archaebacterium Pyrococcus furiosus (PfTrx) as a novel scaffold meeting all the above criteria. PfTrx is a highly thermostable and protease-resistant scaffold with a strong (poly)peptide solubilisation capacity. Anti-PfTrx antibodies did not cross-react with mouse, nor human thioredoxin. Untagged PfTrx bearing a previously identified HPV16-L2 peptide epitope was obtained in a >90% pure form with a one-step thermal purification procedure and effectively elicited the production of neutralizing anti-HPV antibodies. We thus propose PfTrx as a superior, general-purpose scaffold for the construction of safe, stable, and low-cost peptide immunogens.
authors.affiliation: 1] Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy [2].
authors.forename: Elena
authors.initials: E
authors.lastname: Canali
authors.affiliation: 1] Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy [2].
authors.forename: Angelo
authors.initials: A
authors.lastname: Bolchi
authors.affiliation: Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy.
authors.forename: Gloria
authors.initials: G
authors.lastname: Spagnoli
authors.affiliation: German Cancer Research Center, Heidelberg, Germany.
authors.forename: Hanna
authors.initials: H
authors.lastname: Seitz
authors.affiliation: German Cancer Research Center, Heidelberg, Germany.
authors.forename: Ivonne
authors.initials: I
authors.lastname: Rubio
authors.affiliation: C.I.M. Laboratory, Technopole Parma, University of Parma, Italy.
authors.forename: Thelma A
authors.initials: TA
authors.lastname: Pertinhez
authors.affiliation: German Cancer Research Center, Heidelberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Müller
authors.affiliation: Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Ottonello
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008670:Metals / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000737:chemistry / Q000276:immunology; D027383:Papillomaviridae / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016415:Sequence Alignment; D012995:Solubility; D013879:Thioredoxins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D000906:Antibodies; D000956:Antigens, Viral; D001426:Bacterial Proteins; D036022:Capsid Proteins; D000939:Epitopes; C063316:L2 protein, Human papillomavirus type 16; D008670:Metals; D009856:Oncogene Proteins, Viral; D010455:Peptides; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins
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