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Naslov:Recombinant glycoproteins resembling carbohydrate-specific IgE epitopes from plants, venoms and mites
Avtorji:ID Gattinger, Pia (Avtor)
ID Mittermann, Irene (Avtor)
ID Lupinek, Christian (Avtor)
ID Hofer, Gerhard (Avtor)
ID Keller, Walter (Avtor)
ID Bidovec, Urška, Klinika Golnik (Avtor)
ID Korošec, Peter, Klinika Golnik (Avtor)
ID Koessler, Christine (Avtor)
ID Novak, Natalija (Avtor)
ID Valenta, Rudolf (Avtor)
Datoteke:.pdf PDF - Predstavitvena datoteka, prenos (2,07 MB)
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URL URL - Izvorni URL, za dostop obiščite https://www.ebiomedicine.com/article/S2352-3964(18)30573-5/fulltext
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo UKPBAG - Univerzitetna klinika za pljučne bolezni in alergijo Golnik
Povzetek:Background: N-linked glycans present in venoms, pollen and mites are recognized by IgE antibodies from >20% of allergic patients but have low or no allergenic activity. Objectives: To engineer recombinant glycoproteins resembling carbohydrate-specific IgE epitopes from venoms, pollen and mites which can discriminate carbohydrate-specific IgE from allergenic, peptide-specific IgE. Methods: One or two N-glycosylation sites were engineered into the N-terminus of the non-allergenic protein horse heart myoglobin (HHM) using synthetic gene technology. HHM 1 and HHM 2 containing one or two N-glycosylation sites were expressed in baculovirus-infected High-FiveTM insect cells and a non-glycosylated version (HHM 0) was obtained by mutating the glycosylation motif. Recombinant HHM proteins were analyzed regarding fold and aggregation by circular dichroism and gel filtration, respectively. IgE reactivity was assessed by ELISA, immunoblotting and quantitative ImmunoCAP measurements. IgE inhibition assays were performed to study cross-reactivity with venom, plant and mite-derived carbohydrate IgE epitopes. Results: HHM-glycovariants were expressed and purified from insect cells as monomeric and folded proteins. The HHM-glycovariants exhibited strictly carbohydrate-specific IgE reactivity, designed to quantify carbohydrate specific IgE and resembled IgE epitopes of pollen, venom and mite-derived carbohydrates. IgE-reactivity and inhibition experiments established a hierarchy of plant glcyoallergens (nPhl p 4 > nCyn d 1 > nPla a 2 > nJug r 2 > nCup a 1 > nCry j 1) indicating a hitherto unknown heterogeneity of carbohydrate IgE epitopes in plants which were completely represented by HHM 2. Conclusion: Defined recombinant HHM-glycoproteins resembling carbohydrate-specific IgE epitopes from plants, venoms and mites were engineered which made it possible to discriminate carbohydrate- from peptide-specific IgE reactivity.
Ključne besede:allergy and immunology, allergens, glycoproteins, molecular diagnostic technique, recombinant glycoproteins, molecular allergology, component-resolved diagnosis
Leto izida:2019
Status objave pri reviji:Objavljeno
Verzija članka:Založnikova različica članka
Založnik:Elsevier
Št. strani:str. 33-43
Številčenje:Vol. 39
Izvor:Nizozemska
UDK:616-097
ISSN pri članku:2352-3964
DOI:10.1016/j.ebiom.2018.12.002 Novo okno
COBISS.SI-ID:2048486001 Novo okno
Avtorske pravice:© 2018 The Authors
Opomba:Soavtorji: Irene Mittermann, Christian Lupinek, Gerhard Hofer, Walter Keller, Urska Bidovec Stojkovic, Peter Korosec, Christine Koessler, Natalija Novak, Rudolf Valenta; Nasl. z nasl. zaslona; Opis vira z dne 25. 4. 2019;
Datum objave v DiRROS:16.10.2020
Število ogledov:918
Število prenosov:625
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Gradivo je del revije

Naslov:EBioMedicine
Založnik:Elsevier
ISSN:2352-3964
COBISS.SI-ID:523063577 Novo okno

Licence

Licenca:CC BY-NC-ND 4.0, Creative Commons Priznanje avtorstva-Nekomercialno-Brez predelav 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by-nc-nd/4.0/deed.sl
Opis:Najbolj omejujoča licenca Creative Commons. Uporabniki lahko prenesejo in delijo delo v nekomercialne namene in ga ne smejo uporabiti za nobene druge namene.
Začetek licenciranja:03.12.2018

Sekundarni jezik

Jezik:Ni določen
Ključne besede:alergija in imunologija, alergeni, glikoproteini, molekularne diagnostične tehnike, rekombinantni glikoproteini, molekularna alergologija, molekularna diagnostika


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