Open AccessHigh‐yield production of a human monoclonal IgG by rhizosecretion in hydroponic tobacco cultures
Author(s) -
Madeira Luisa M.,
Szeto Tim H.,
Henquet Maurice,
Raven Nicole,
Runions John,
Huddleston Jon,
Garrard Ian,
Drake Pascal M.W.,
Ma Julian KC.
Publication year - 2016
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12407
Subject(s) - chinese hamster ovary cell , recombinant dna , monoclonal antibody , biology , in vitro , affinity chromatography , antibody , biochemistry , microbiology and biotechnology , gene , enzyme , receptor , immunology
Summary 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 IgG 1 monoclonal antibody ( mA b) 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, mA b yields in hydroponic medium reached 46 μg/mL in 1 week, the highest figure reported for a recombinant mA b in a plant secretion‐based system to date. The rhizosecretome was determined to contain 104 proteins, with the mA b 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 mA b 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.