Purification by Strep-Tactin Affinity Chromatography of a Delete Envelope gp51 Protein of Bovine Leukaemia Virus Expressed in Sf21 Insect Cells |
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Authors: | Antonio De Giuseppe Katia Forti Francesco Feliziani Giulio Severi Monica Cagiola |
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Institution: | (1) Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, G. Salvemini 1, 06126 Perugia, Italy |
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Abstract: | Bovine leukaemia virus (BLV) causes disease in cattle and it is related to human T lymphotrofic viruses HTLV-1 and HTLV-2.
The objective of this study was to express and purify deleted and stable forms of the gp51 envelope glycoprotein of BLV using
a baculovirus system. Two forms of the gp51 were synthesised: one comprised the gp51 N-terminal 174 amino acids and a single
6xHis tag (∆175-268gp51-His) and the second form contained the same amino acid sequence and a C-terminal Strep-tag II in addition to the 6xHis
tag (∆175-268gp51-STH). The two proteins were expressed and purified by immobilized metal-affinity chromatography (IMAC) or by Strep-Tactin
column. The Strep-Tactin technology was more efficient than IMAC method and achieved a high pure recombinant deleted gp51.
In addition the ∆175-268gp51-STH protein was further concentrated by IMAC. This purified antigen could be used for the isolation of immunoreactive
molecules and to develop a competitive ELISA test. |
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