Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens
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2015
Authors
Popović, Milica M.Prodanović, Radivoje
Ostafe, Raluca
Schillberg, Stefan
Fischer, Rainer
Gavrović-Jankulović, Marija
Article (Published version)
Metadata
Show full item recordAbstract
High-throughput characterization of allergens relies often on phage display technique which is subject to the limitations of a prokaryotic expression system. Substituting the phage display platform with a yeast surface display could lead to fast immunological characterization of allergens with complex structures. Our objective was to evaluate the potential of yeast surface display for characterization of plant-derived food allergens. The coding sequence of mature actinidin (Act d 1) was cloned into pCTCON2 surface display vector. Flow cytometry was used to confirm localization of recombinant Act d 1 on the surface of yeast cells using rabbit polyclonal antisera IgG and IgE from sera of kiwifruit-allergic individuals. Immunological (dot blot, immunoblot ELISA and ELISA inhibition), biochemical (enzymatic activity in gel) and biological (basophil activation) characterization of Act d 1 after solubilization from the yeast cell confirmed that recombinant Act d 1 produced on the surface of ...yeast cell is similar to its natural counterpart isolated from green kiwifruit. Yeast surface display is a potent technique that enables fast immunochemical characterization of allergens in situ without the need for protein purification and offers an alternative that could lead to improvement of standard immunodiagnostic and immunotherapeutic approaches.
Keywords:
Actinidin / Allergen characterization / Cysteine protease / Kiwifruit / Yeast surface displaySource:
Immunologic Research, 2015, 61, 3, 230-239Publisher:
- Humana Press Inc, Totowa
Funding / projects:
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-MESTD-Basic Research (BR or ON)-172049)
- FEBS Collaborative and Experimental Scholarship for Central and Eastern Europe
DOI: 10.1007/s12026-014-8614-0
ISSN: 0257-277X
PubMed: 25537533
WoS: 000350295900006
Scopus: 2-s2.0-84925518006
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Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Popović, Milica M. AU - Prodanović, Radivoje AU - Ostafe, Raluca AU - Schillberg, Stefan AU - Fischer, Rainer AU - Gavrović-Jankulović, Marija PY - 2015 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1668 AB - High-throughput characterization of allergens relies often on phage display technique which is subject to the limitations of a prokaryotic expression system. Substituting the phage display platform with a yeast surface display could lead to fast immunological characterization of allergens with complex structures. Our objective was to evaluate the potential of yeast surface display for characterization of plant-derived food allergens. The coding sequence of mature actinidin (Act d 1) was cloned into pCTCON2 surface display vector. Flow cytometry was used to confirm localization of recombinant Act d 1 on the surface of yeast cells using rabbit polyclonal antisera IgG and IgE from sera of kiwifruit-allergic individuals. Immunological (dot blot, immunoblot ELISA and ELISA inhibition), biochemical (enzymatic activity in gel) and biological (basophil activation) characterization of Act d 1 after solubilization from the yeast cell confirmed that recombinant Act d 1 produced on the surface of yeast cell is similar to its natural counterpart isolated from green kiwifruit. Yeast surface display is a potent technique that enables fast immunochemical characterization of allergens in situ without the need for protein purification and offers an alternative that could lead to improvement of standard immunodiagnostic and immunotherapeutic approaches. PB - Humana Press Inc, Totowa T2 - Immunologic Research T1 - Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens VL - 61 IS - 3 SP - 230 EP - 239 DO - 10.1007/s12026-014-8614-0 ER -
@article{ author = "Popović, Milica M. and Prodanović, Radivoje and Ostafe, Raluca and Schillberg, Stefan and Fischer, Rainer and Gavrović-Jankulović, Marija", year = "2015", abstract = "High-throughput characterization of allergens relies often on phage display technique which is subject to the limitations of a prokaryotic expression system. Substituting the phage display platform with a yeast surface display could lead to fast immunological characterization of allergens with complex structures. Our objective was to evaluate the potential of yeast surface display for characterization of plant-derived food allergens. The coding sequence of mature actinidin (Act d 1) was cloned into pCTCON2 surface display vector. Flow cytometry was used to confirm localization of recombinant Act d 1 on the surface of yeast cells using rabbit polyclonal antisera IgG and IgE from sera of kiwifruit-allergic individuals. Immunological (dot blot, immunoblot ELISA and ELISA inhibition), biochemical (enzymatic activity in gel) and biological (basophil activation) characterization of Act d 1 after solubilization from the yeast cell confirmed that recombinant Act d 1 produced on the surface of yeast cell is similar to its natural counterpart isolated from green kiwifruit. Yeast surface display is a potent technique that enables fast immunochemical characterization of allergens in situ without the need for protein purification and offers an alternative that could lead to improvement of standard immunodiagnostic and immunotherapeutic approaches.", publisher = "Humana Press Inc, Totowa", journal = "Immunologic Research", title = "Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens", volume = "61", number = "3", pages = "230-239", doi = "10.1007/s12026-014-8614-0" }
Popović, M. M., Prodanović, R., Ostafe, R., Schillberg, S., Fischer, R.,& Gavrović-Jankulović, M.. (2015). Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens. in Immunologic Research Humana Press Inc, Totowa., 61(3), 230-239. https://doi.org/10.1007/s12026-014-8614-0
Popović MM, Prodanović R, Ostafe R, Schillberg S, Fischer R, Gavrović-Jankulović M. Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens. in Immunologic Research. 2015;61(3):230-239. doi:10.1007/s12026-014-8614-0 .
Popović, Milica M., Prodanović, Radivoje, Ostafe, Raluca, Schillberg, Stefan, Fischer, Rainer, Gavrović-Jankulović, Marija, "Yeast surface display is a novel tool for the rapid immunological characterization of plant-derived food allergens" in Immunologic Research, 61, no. 3 (2015):230-239, https://doi.org/10.1007/s12026-014-8614-0 . .