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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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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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Abbreviations

BLV:

Bovine leukaemia virus

HTLV:

Human T-cell leukaemia virus

FLK:

Foetal lamb kidney

EBL:

Enzootic bovine leucosis

IMAC:

Metal-affinity chromatography

PBS:

Phosphate buffer saline

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Acknowledgments

The authors thank Ms Tamsin Anne Hickson for the English language revisions.

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Authors and Affiliations

  1. Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, G. Salvemini 1, 06126, Perugia, Italy

    Antonio De Giuseppe, Katia Forti, Francesco Feliziani, Giulio Severi & Monica Cagiola

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  1. Antonio De Giuseppe
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  2. Katia Forti
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  3. Francesco Feliziani
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  4. Giulio Severi
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  5. Monica Cagiola
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Correspondence to Antonio De Giuseppe.

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De Giuseppe, A., Forti, K., Feliziani, F. et al. Purification by Strep-Tactin Affinity Chromatography of a Delete Envelope gp51 Protein of Bovine Leukaemia Virus Expressed in Sf21 Insect Cells. Protein J 29, 153–160 (2010). https://doi.org/10.1007/s10930-010-9228-6

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  • DOI: https://doi.org/10.1007/s10930-010-9228-6

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