title: Characterization of VRC01, a potent and broadly neutralizing anti-HIV mAb, produced in transiently and stably transformed tobacco.
abstract: The proposed clinical trial in Africa of VRC01, a potent broadly neutralizing antibody (bNAb) capable of neutralizing 91% of known HIV-1 isolates, raises concerns about testing a treatment which will be too expensive to be accessible by the most important target population, the poor in under-developed regions such as sub-Saharan Africa. Here, we report the expression of VRC01 in plants as an economic alternative to conventional mammalian-cell-based production platforms. The heavy and light chain genes of VRC01 were cloned onto a single vector, pTRAk.2, which was transformed into Nicotiana benthamiana or Nicotiana tabacum using transient and stable expression production systems respectively. VRC01 has been successfully expressed transiently in plants with expression level of approximately 80 mg antibody/kg; stable transgenic lines expressing up to 100 mg antibody/kg were also obtained. Plant-produced VRC01 from both systems showed a largely homogeneous N-glycosylation profile with a single dominant glycoform. The binding kinetics to gp120 IIIB (approximately 1 nM), neutralization of HIV-1 BaL or a panel of 10 VRC01-sensitive HIV-1 Env pseudoviruses of VRC01 produced in transient and stable plants were also consistent with VRC01 from HEK cells.
authors.affiliation: Molecular Immunology Unit, Infection and Immunity Research Centre, St. George's University of London, London, UK.
authors.forename: Audrey Y-H
authors.initials: AY
authors.lastname: Teh
authors.forename: Daniel
authors.initials: D
authors.lastname: Maresch
authors.forename: Katja
authors.initials: K
authors.lastname: Klein
authors.forename: Julian K-C
authors.initials: JK
authors.lastname: Ma
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D057134:Antibodies, Neutralizing / Q000096:biosynthesis / Q000302:isolation & purification; D000080908:Broadly Neutralizing Antibodies; D002461:Cell Line, Transformed; D015870:Gene Expression; D006031:Glycosylation; D015483:HIV Antibodies / Q000096:biosynthesis / Q000302:isolation & purification*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011485:Protein Binding; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000080908:Broadly Neutralizing Antibodies; D015483:HIV Antibodies; C578595:VRC01 monoclonal antibody
keywords: HIV-1 broadly neutralizing antibody; VRC01; pTRAk.2; stable transgenics; transient expression