Madeira, LM; Szeto, TH; Ma, JK; Drake, PM
(2016)
Rhizosecretion improves the production of Cyanovirin-N in Nicotiana tabacum through simplified downstream processing.
Biotechnology Journal, 11 (7).
pp. 910-919.
ISSN 1860-7314
https://doi.org/10.1002/biot.201500371
SGUL Authors: Drake, Pascal
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Abstract
Rhizosecretion has many advantages for the production of recombinant pharmaceuticals, notably facile downstream processing from hydroponic medium. The aim of this study was to increase yields of the HIV microbicide candidate, Cyanovirin-N (CV-N), obtained using this production platform and to develop a simplified methodology for its downstream processing from hydroponic medium. Placing hydroponic cultures on an orbital shaker more than doubled the concentration of CV-N in the hydroponic medium compared to plants which remained stationary, reaching a maximum of approximately 20μg/ml in one week, which is more than 3 times higher than previously reported yields. The protein composition of the hydroponic medium, the rhizosecretome, was characterised in plants cultured with or without the plant growth regulator alpha-napthaleneacetic acid by LC-ESI-MS/MS, and CV-N was the most abundant protein. The issue of large volumes in the rhizosecretion system was addressed by using ion exchange chromatography to concentrate CV-N and partially remove impurities. The semi-purified CV-N was demonstrated to bind to HIV gp120 in an ELISA and to neutralise HIVBa-L with an IC50 of 6nM in a cell-based assay. Rhizosecretion is therefore a practicable and inexpensive method for the production of functional CV-N.
Item Type: | Article | |||||||||
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Additional Information: | This is the peer reviewed version of the following article: Madeira, L. M., Szeto, T. H., Ma, J. K.-C. and Drake, P. M.W. (2016), Rhizosecretion improves the production of Cyanovirin-N in Nicotiana tabacum through simplified downstream processing. Biotechnology Journal, 11: 910–919, which has been published in final form at http://dx.doi.org/10.1002/biot.201500371. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. | |||||||||
Keywords: | Cyanovirin-N, HIV microbicide, Hydroponic medium, Molecular pharming, Rhizosecretion, Biotechnology, 1002 Environmental Biotechnology, 1003 Industrial Biotechnology, 1004 Medical Biotechnology | |||||||||
SGUL Research Institute / Research Centre: | Academic Structure > Infection and Immunity Research Institute (INII) | |||||||||
Journal or Publication Title: | Biotechnology Journal | |||||||||
ISSN: | 1860-7314 | |||||||||
Language: | ENG | |||||||||
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Publisher License: | Publisher's own licence | |||||||||
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PubMed ID: | 26901579 | |||||||||
Go to PubMed abstract | ||||||||||
URI: | https://openaccess.sgul.ac.uk/id/eprint/107780 | |||||||||
Publisher's version: | https://doi.org/10.1002/biot.201500371 |
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