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Evaluation of thermo-stability of bluetongue virus recombinant VP7 antigen in indirect ELISA

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Abstract

This study shows the thermo-stability of lyophilized and purified recombinant VP7 bluetongue virus (BTV) protein in the presence of two sugar stabilizers (trehalose and mannitol) at different temperature. Truncated VP7 protein purified by nickel affinity column was lyophilized in the presence of trehalose and mannitol at 60 mM final concentration and then exposed to different temperature like 4, 25, 37 and 45 °C for various periods like 5 months, 7 weeks, 7 days and 48 h, respectively. After thermal treatment, the reactivity of the protein was evaluated in indirect ELISA. At 4 and 25 °C, the protein was stable up to 5 months and 7 weeks, respectively, irrespective of stabilizers used. At 37 °C, it was stable up to 3 days with both the stabilizers, after which it lost its stability and reactivity. At 45 °C, the protein was stable up to 30 and 24 h with trehalose and mannitol stabilizers, respectively. Both stabilizers found suitable for stability of the protein. However, trehalose appeared to have better stabilizing effect, particularly at higher temperatures than the mannitol. Trehalose could be used as stabilizer for freeze-drying the recombinant VP7 protein if an indirect ELISA kit based on the purified rVP7 protein is supplied to different laboratories of the country for detection of BTV antibody in sheep.

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Acknowledgments

The authors thank the Director, Indian Veterinary Research Institute for providing necessary facilities to carry out this work and the staff of Tissue Culture Laboratory. This is first author’s M.V.Sc thesis work which has been carried out under the financial assistance of All India Network project on Bluetongue (AINP-BT), Indian Council of Agricultural Research (ICAR), New Delhi, India.

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Authors and Affiliations

  1. Division of Virology, Indian Veterinary Research Institute, Mukteswar Campus, Dist. Nainital, Mukteswar, 263 138, Uttarakhand, India

    Gnanavel Venkatesan, Sanchay Kumar Biswas, Veerakyathappa Bhanuprakash, Raj Kumar Singh & Bimelendu Mondal

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  1. Gnanavel Venkatesan
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  2. Sanchay Kumar Biswas
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  3. Veerakyathappa Bhanuprakash
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  4. Raj Kumar Singh
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  5. Bimelendu Mondal
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Correspondence to Gnanavel Venkatesan.

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Venkatesan, G., Biswas, S.K., Bhanuprakash, V. et al. Evaluation of thermo-stability of bluetongue virus recombinant VP7 antigen in indirect ELISA. VirusDis. 26, 19–26 (2015). https://doi.org/10.1007/s13337-014-0244-6

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  • DOI: https://doi.org/10.1007/s13337-014-0244-6

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