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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Recombinant expression of trypanosome surface glycoproteins in Pichia pastoris for the diagnosis of Trypanosoma evansi infection
Veterinary Parasitology, Volume 197, No. 3-4, Year 2013
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Description
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 Man5GlcNAc2 N-glycosylation which resembles the predominant Man9-5GlcNAc2 oligomannose structures in T. brucei. The secreted recombinant antigens were affinity purified with yields of up to 10mg and 20mg per liter cell culture of rISG 7529-465-E and rRoTat 1.223-385-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 1.223-385-H but not of rISG 7529-465-E was confirmed with sera of naturally infected and control dromedary camels. The results suggest that rRoTat 1.223-385-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. © 2013 Elsevier B.V.
Authors & Co-Authors
Rogé, S.
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Belgium, Antwerpen
Universiteit Antwerpen
van Reet, Nick
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Odiwuor, Samwel O.
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Kenya, Thika
Mount Kenya University
Tran, Thao
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Schildermans, K.
Belgium, Antwerpen
Universiteit Antwerpen
Vandamme, S.
Belgium, Antwerpen
Universiteit Antwerpen
Vandenberghe, Isabel
Belgium, Ghent
Universiteit Gent
Vervecken, W.
Belgium, Ghent
Vlaams Instituut Voor Biotechnologie
Belgium, Ghent
Universiteit Gent
Gillingwater, Kirsten
Switzerland, Allschwil
Swiss Tropical and Public Health Institute Swiss Tph
Claes, Filip
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Devreese, Bart V.
Belgium, Ghent
Universiteit Gent
Guisez, Yves
Belgium, Antwerpen
Universiteit Antwerpen
Büscher, Philippe
Belgium, Antwerpen
Prins Leopold Instituut Voor Tropische Geneeskunde
Statistics
Citations: 28
Authors: 13
Affiliations: 6
Identifiers
Doi:
10.1016/j.vetpar.2013.05.009
ISSN:
03044017
Research Areas
Genetics And Genomics