title: Glycan modulation and sulfoengineering of anti-HIV-1 monoclonal antibody PG9 in plants.
abstract: Broadly neutralizing anti-HIV-1 monoclonal antibodies, such as PG9, and its derivative RSH hold great promise in AIDS therapy and prevention. An important feature related to the exceptional efficacy of PG9 and RSH is the presence of sulfated tyrosine residues in their antigen-binding regions. To maximize antibody functionalities, we have now produced glycan-optimized, fucose-free versions of PG9 and RSH in Nicotiana benthamiana. Both antibodies were efficiently sulfated in planta on coexpression of an engineered human tyrosylprotein sulfotransferase, resulting in antigen-binding and virus neutralization activities equivalent to PG9 synthesized by mammalian cells ((CHO)PG9). Based on the controlled production of both sulfated and nonsulfated variants in plants, we could unequivocally prove that tyrosine sulfation is critical for the potency of PG9 and RSH. Moreover, the fucose-free antibodies generated in N. benthamiana are capable of inducing antibody-dependent cellular cytotoxicity, an activity not observed for (CHO)PG9. Thus, tailoring of the antigen-binding site combined with glycan modulation and sulfoengineering yielded plant-produced anti-HIV-1 antibodies with effector functions superior to PG9 made in CHO cells.
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Andreas
authors.initials: A
authors.lastname: Loos
authors.affiliation: Division of Infectious Diseases, University of California, Irvine, CA 92697;
authors.forename: Johannes S
authors.initials: JS
authors.lastname: Gach
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Thomas
authors.initials: T
authors.lastname: Hackl
authors.affiliation: Department of Chemistry, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Daniel
authors.initials: D
authors.lastname: Maresch
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Theresa
authors.initials: T
authors.lastname: Henkel
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Andreas
authors.initials: A
authors.lastname: Porodko
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Duc
authors.initials: D
authors.lastname: Bui-Minh
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sommeregger
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria.
authors.forename: Gordana
authors.initials: G
authors.lastname: Wozniak-Knopp
authors.affiliation: Division of Infectious Diseases, University of California, Irvine, CA 92697;
authors.forename: Donald N
authors.initials: DN
authors.lastname: Forthal
authors.affiliation: Department of Chemistry, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria;
authors.forename: Friedrich
authors.initials: F
authors.lastname: Altmann
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria; herta.steinkellner@boku.ac.at lukas.mach@boku.ac.at.
authors.forename: Herta
authors.initials: H
authors.lastname: Steinkellner
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, A-1190 Vienna, Austria; herta.steinkellner@boku.ac.at lukas.mach@boku.ac.at.
authors.forename: Lukas
authors.initials: L
authors.lastname: Mach
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D015483:HIV Antibodies / Q000096:biosynthesis; D015497:HIV-1; D006801:Humans; D060847:Metabolic Engineering / Q000379:methods*; D011134:Polysaccharides / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D011134:Polysaccharides; D011994:Recombinant Proteins
keywords: antibody; biopharmaceutical; glycosylation; plant; sulfation