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.
TY - 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
UR - Kon_2814
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",
url = "Kon_2814"
}
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
Kon_2814
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
Kon_2814 .
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 .,
Kon_2814 .
