Hiszczyńska-Sawicka Elzbieta, Brillowska-Dabrowska Anna, Dabrowski Sławomir, Pietkiewicz Halina, Myjak Przemysław, Kur Józef
Department of Microbiology, Technical University of Gdańsk, ul. Narutowicza 11/12, 80-952 Gdańsk, Poland.
Protein Expr Purif. 2003 Jan;27(1):150-7. doi: 10.1016/s1046-5928(02)00593-4.
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 Ni(2+)-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.
本报告描述了一种简单、高效且可重复的方法，用于获得大量高纯度的重组弓形虫抗原，这些抗原可用于诊断应用。所获得的在大肠杆菌中表达的弓形虫SAG1、GRA1和GRA7抗原（作为融合蛋白），在N端和C端含有多组氨酸标签，这使得可以通过在Ni(2+)-IDA-琼脂糖柱上进行金属亲和层析进行单步分离。以酶联免疫吸附测定（ELISA）形式测试了重组抗原的免疫反应性，以评估其在弓形虫感染血清学诊断中的潜在应用。
