title: Monoclonal Antibody Purification (Nicotiana benthamianaPlants).
abstract: Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matobaet al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue ofNicotiana benthamiana(N. benthamiana). To overcome the issues of genetic instability and limited infectivity of recombinant viruses,Agrobacterium-mediated delivery of "deconstructed" virus vectors has become the mainstay for the production of large and/or multicomponent proteins, such as immunoglobulin (Ig)G monoclonal antibodies (mAbs). Here, we describe a method of producing human IgG mAbs inN. benthamianausing the tobamoviral replicon vector magnICON®. The vector can express up to a few hundred mg of a mAb per kg of leaf material in 7 days. A representative case for the broadly neutralizing anti-HIV and anti-influenza mAbs, VRC01 and CR6261 respectively, is shown (Hamorskyet al., 2013). Leaf tissue is homogenized and the extract is clarified by filtration and centrifugation. The mAb is purified by fast protein liquid chromatography (FPLC) using Protein A affinity and Phenyl HP hydrophobic interection resins.
authors.affiliation: Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Husk
authors.affiliation: Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA.
authors.forename: Krystal Teasley
authors.initials: KT
authors.lastname: Hamorsky
authors.affiliation: Department of Pharmacology and Toxicology, and Owensboro Cancer Research Program at James Graham Brown Cancer Center, University of Louisville School of Medicine, Owensboro, USA.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Matoba