Cosecretion of protease inhibitor stabilizes antibodies produced by plant roots.
A plant-based system for continuous production of monoclonal antibodies based on the secretion of immunoglobulin complexes from plant roots into a hydroponic medium (rhizosecretion) was engineered to produce high levels of single-chain and full-size immunoglobulins. Replacing the original signal peptides of monoclonal antibodies with a plant-derived calreticulin signal increased the levels of antibody yield 2-fold. Cosecretion of Bowman-Birk Ser protease inhibitor reduced degradation of the immunoglobulin complexes in the default secretion pathway and further increased antibody production to 36.4 microg/g root dry weight per day for single-chain IgG1 and 21.8 microg/g root dry weight per day for full-size IgG4 antibodies. These results suggest that constitutive cosecretion of a protease inhibitor combined with the use of the plant signal peptide and the antibiotic marker-free transformation system offers a novel strategy to achieve high yields of complex therapeutic proteins secreted from plant roots.
Komarnytsky
Slavko
S
Biotech Center, Rutgers University, New Brunswick, New Jersey 08901, USA. komar@aesop.rutgers.edu
Borisjuk
Nikolai
N
Yakoby
Nir
N
Garvey
Alison
A
Raskin
Ilya
I
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D037282:Calreticulin / Q000378:metabolism; D003470:Culture Media; D006801:Humans; D018527:Hydroponics; D007074:Immunoglobulin G / Q000096:biosynthesis; D018517:Plant Roots / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000235:genetics
D000911:Antibodies, Monoclonal; D037282:Calreticulin; D003470:Culture Media; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D014358:Trypsin Inhibitor, Bowman-Birk Soybean