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Recombinant antigens expressed in Pichia pastoris for the diagnosis of sleeping sickness caused by Trypanosoma brucei gambiense

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Show simple item record Rogé, S. en_US Van Nieuwenhove, L. en_US Meul, M. en_US Heykers, A. en_US Brouwer de Koning, A. en_US Bebronne, N. en_US Guisez, Y. en_US Büscher, P. en_US 2014-09-25T13:39:57Z 2014-09-25T13:39:57Z 2014 en_US
dc.identifier.issn 1935-2727 en_US
dc.identifier.doi en_US
dc.identifier.other ITG-B1A; ITG-B2B; ITG-B3B; ITG-B4B; ITG-B5B; ITG-B6A; ITG-BLA; DBM; U-PARDIA; JIF; DOI; FTA; OAJ; E-only; Abstract; UPD56 en_US
dc.description.abstract BACKGROUND: Screening tests for gambiense sleeping sickness, such as the CATT/T. b. gambiense and a recently developed lateral flow tests, are hitherto based on native variant surface glycoproteins (VSGs), namely LiTat 1.3 and LiTat 1.5, purified from highly virulent trypanosome strains grown in rodents. METHODOLOGY/PRINCIPAL FINDINGS: We have expressed SUMO (small ubiquitin-like modifier) fusion proteins of the immunogenic N-terminal part of these antigens in the yeast Pichia pastoris. The secreted recombinant proteins were affinity purified with yields up to 10 mg per liter cell culture. CONCLUSIONS/SIGNIFICANCE: The diagnostic potential of each separate antigen and a mixture of both antigens was confirmed in ELISA on sera from 88 HAT patients and 74 endemic non-HAT controls. Replacement of native antigens in the screening tests for sleeping sickness by recombinant proteins will eliminate both the infection risk for the laboratory staff during antigen production and the need for laboratory animals. Upscaling production of recombinant antigens, e.g. in biofermentors, is straightforward thus leading to improved standardisation of antigen production and reduced production costs, which on their turn will increase the availability and affordability of the diagnostic tests needed for the elimination of gambiense HAT. en_US
dc.language English en_US
dc.subject Protozoal diseases en_US
dc.subject Sleeping sickness en_US
dc.subject Trypanosoma brucei gambiense en_US
dc.subject Vectors en_US
dc.subject Tsetse flies en_US
dc.subject Glossina morsitans morsitans en_US
dc.subject Diagnosis en_US
dc.subject Antigen detection en_US
dc.subject Protein expression en_US
dc.subject Recombinant antigen en_US
dc.subject Yeasts en_US
dc.subject Pichia pastoris en_US
dc.subject Confirmation en_US
dc.subject ELISA en_US
dc.subject Laboratory security en_US
dc.subject Laboratory animals en_US
dc.subject Laboratory techniques and procedures en_US
dc.title Recombinant antigens expressed in Pichia pastoris for the diagnosis of sleeping sickness caused by Trypanosoma brucei gambiense en_US
dc.type Article-E en_US
dc.citation.issue 7 en_US
dc.citation.jtitle PLoS Neglected Tropical Diseases en_US
dc.citation.volume 8 en_US
dc.citation.pages e3006 en_US
dc.identifier.pmid en_US
dc.citation.jabbreviation PLoS Negl Trop Dis en_US

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