Comparison of microbial and transient expression (tobacco plants and plant-cell packs) for the production and purification of the anti-cancer mistl…
Authors of this article are:
Gengenbach BB, Keil LL, Opdensteinen P, Müschen CR, Melmer G, Lentzen H, Bührmann J, Buyel JF.
A summary of the article is shown below:
Cancer is the leading cause of death in industrialized countries. Cancer therapy often involves monoclonal antibodies or small-molecule drugs, but carbohydrate-binding lectins such as mistletoe (Viscum album) viscumin offer a potential alternative treatment strategy. Viscumin is toxic in mammalian cells, ruling them out as an efficient production system, and it forms inclusion bodies in Escherichia coli such that purification requires complex and lengthy refolding steps. We therefore investigated the transient expression of viscumin in intact Nicotiana benthamiana plants and N. tabacum BY-2 plant-cell packs (PCPs), comparing a full-length viscumin gene construct to separate constructs for the A and B chains. As determined by capillary electrophoresis the maximum yield of purified heterodimeric viscumin in N. benthamiana was ~7 mg kg-1 fresh biomass with the full-length construct. The yield was about 50% higher in PCPs but reduced 10-fold when co-expressing A and B chains as individual polypeptides. Using a single-step lactosyl-Sepharose affinity resin, we purified viscumin to ~54%. The absence of refolding steps resulted in estimated cost savings of more than 80% when transient expression in tobacco was compared to E. coli. Furthermore, the plant-derived product was ~3-fold more toxic than the bacterially-produced counterpart. We conclude that plants offer a suitable alternative for the production of complex biopharmaceutical proteins that are toxic to mammalian cells and that form inclusion bodies in bacteria. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.
Check out the article’s website on Pubmed for more information:
This article is a good source of information and a good way to become familiar with topics such as: Cancer therapy; Design of experiments; Molecular farming; Plant-cell pack; Recombinant viscumin; Transient protein expression.
