Title
Recombinant expression of trypanosome surface glycoproteins in **Pichia pastoris** for the diagnosis of **Trypanosoma evansi** infection Recombinant expression of trypanosome surface glycoproteins in **Pichia pastoris** for the diagnosis of **Trypanosoma evansi** infection
Author
Faculty/Department
Faculty of Sciences. Biology
Publication type
article
Publication
Amsterdam ,
Subject
Veterinary medicine
Human medicine
Source (journal)
Veterinary parasitology. - Amsterdam
Volume/pages
197(2013) :3-4 , p. 571-579
ISSN
0304-4017
ISI
000328010500021
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Serodiagnosis of surra, which causes vast economic losses in livestock, is still based on native antigens purified from bloodstream form Trypanosoma (T.) evansi grown in rodents. To avoid the use of laboratory rodents in antigen preparation we expressed fragments of the invariant surface glycoprotein (ISG) 75, cloned from T. brucei gambiense cDNA, and the variant surface glycoprotein (VSG) RoTat 1.2, cloned from T. evansi gDNA, recombinantly in Pichia (P.) pastoris. The M5 strain of this yeast has an engineered N-glycosylation pathway resulting in homogenous Man(5)GlcNAc(2) N-glycosylation which resembles the predominant Man(9-5)GlcNAc(2) oligomannose structures in T. brucei. The secreted recombinant antigens were affinity purified with yields of up to 10 mg and 20 mg per liter cell culture of rISG 75(29-465-E) and rRoTat 1.2(23-385-H) respectively. In ELISA, both recombinant proteins discriminated between pre-immune and immune serum samples of 25 goats experimentally infected with T. evansi. The diagnostic potential of rRoTat 1.2(23-385-H) but not of rISG 75(29-465-E) was confirmed with sera of naturally infected and control dromedary camels. The results suggest that rRoTat 1.2(23-385-H) expressed in P. pastoris requires further evaluation before it could replace native RoTat 1.2 VSG for serodiagnosis of surra, thus eliminating the use of laboratory animals for antigen production. (C) 2013 Elsevier B.V. All rights reserved.
E-info
https://repository.uantwerpen.be/docman/iruaauth/508708/36c6507.pdf
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