4.2 Article

High yield expression and single-step purification of Toxoplasma gondii SAG1, GRA1, and GRA7 antigens in Escherichia coli

期刊

PROTEIN EXPRESSION AND PURIFICATION
卷 27, 期 1, 页码 150-157

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ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/S1046-5928(02)00593-4

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ELISA; fusion protein; metal-affinity chromatography; serological detection; Toxoplasma gondii antigens; Western blot

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This report describes a simple, highly efficient and reproducible method for obtaining large quantities of highly pure recombinant Toxoplasma gondii antigens, which can be used for diagnostic application. The obtained T gondii SAG1, GRA1, and GRA7 antigens (as fusion proteins), expressed in Escherichia coli, contained polyhistidine tags at the N- and C-ends that allowed single-step isolation by metal-affinity chromatography on Ni2+-IDA-Sepharose columns. The immunoreactivity of the recombinant antigens was tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of T gondii infection. (C) 2002 Elsevier Science (USA). All rights reserved.

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