Format

Send to

Choose Destination

See 1 citation found by title matching your search:

Plant Biotechnol J. 2016 Feb;14(2):615-24. doi: 10.1111/pbi.12407. Epub 2015 Jun 3.

High-yield production of a human monoclonal IgG by rhizosecretion in hydroponic tobacco cultures.

Author information

1
The Hotung Molecular Immunology Unit, Institute for Infection and Immunity, St. George's University of London, London, UK.
2
Plant Research International, Wageningen University and Research Centre, Wageningen, The Netherlands.
3
Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Aachen, Germany.
4
Department of Biological and Medical Sciences - Faculty of Health and Life Sciences, Oxford Brookes University, Oxford, UK.
5
Brunel Institute for Bioengineering, Brunel University, London, UK.

Abstract

Rhizosecretion of recombinant pharmaceuticals from in vitro hydroponic transgenic plant cultures is a simple, low cost, reproducible and controllable production method. Here, we demonstrate the application and adaptation of this manufacturing platform to a human antivitronectin IgG1 monoclonal antibody (mAb) called M12. The rationale for specific growth medium additives was established by phenotypic analysis of root structure and by LC-ESI-MS/MS profiling of the total protein content profile of the hydroponic medium. Through a combination of optimization approaches, mAb yields in hydroponic medium reached 46 μg/mL in 1 week, the highest figure reported for a recombinant mAb in a plant secretion-based system to date. The rhizosecretome was determined to contain 104 proteins, with the mAb heavy and light chains the most abundant. This enabled evaluation of a simple, scalable extraction and purification protocol and demonstration that only minimal processing was necessary prior to protein A affinity chromatography. MALDI-TOF MS revealed that purified mAb contained predominantly complex-type plant N-glycans, in three major glycoforms. The binding of M12 purified from hydroponic medium to vitronectin was comparable to its Chinese hamster ovary (CHO)-derived counterpart. This study demonstrates that in vitro hydroponic cultivation coupled with recombinant protein rhizosecretion can be a practical, low-cost production platform for monoclonal antibodies.

KEYWORDS:

genetically modified plants; monoclonal antibody; recombinant antibody production; rhizosecretion

PMID:
26038982
DOI:
10.1111/pbi.12407
[Indexed for MEDLINE]
Free full text

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center