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Transgenic Res. 2015 Oct;24(5):897-909. doi: 10.1007/s11248-015-9890-8. Epub 2015 Jun 25.

Tobacco seeds as efficient production platform for a biologically active anti-HBsAg monoclonal antibody.

Author information

1
Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba. abel.hernandez@cigb.edu.cu.
2
Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
3
Department of Carbohydrate Chemistry, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba.
4
Department of Plant Systems Biology, VIB, Plant-made Antibodies and Immunogens, Ghent, Belgium.

Abstract

The use of plants as heterologous hosts is one of the most promising technologies for manufacturing valuable recombinant proteins. Plant seeds, in particular, constitute ideal production platforms for long-term applications requiring a steady supply of starting material, as they combine the general advantages of plants as bioreactors with the possibility of biomass storage for long periods in a relatively small volume, thus allowing manufacturers to decouple upstream and downstream processing. In the present work we have used transgenic tobacco seeds to produce large amounts of a functionally active mouse monoclonal antibody against the Hepatitis B Virus surface antigen, fused to a KDEL endoplasmic reticulum retrieval motif, under control of regulatory sequences from common bean (Phaseolus vulgaris) seed storage proteins. The antibody accumulated to levels of 6.5 mg/g of seed in the T3 generation, and was purified by Protein A affinity chromatography combined with SEC-HPLC. N-glycan analysis indicated that, despite the KDEL signal, the seed-derived plantibody bore both high-mannose and complex-type sugars that indicate partial passage through the Golgi compartment, although its performance in the immunoaffinity purification of HBsAg was unaffected. An analysis discussing the industrial feasibility of replacing the currently used tobacco leaf-derived plantibody with this seed-derived variant is also presented.

KEYWORDS:

Hepatitis B; Monoclonal antibody; N-glycan; Phaseolin promoter; Protein expression; Tobacco seeds

PMID:
26109093
DOI:
10.1007/s11248-015-9890-8
[Indexed for MEDLINE]

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