title: Rapid, high-yield production in plants of individualized idiotype vaccines for non-Hodgkin's lymphoma.
abstract: BACKGROUND
  Animal and clinical studies with plant-produced single-chain variable fragment lymphoma vaccines have demonstrated specific immunogenicity and safety. However, the expression levels of such fragments were highly variable and required complex engineering of the linkers. Moreover, the downstream processing could not be built around standard methods like protein A affinity capture.
  
  
  METHODS
  We report a novel vaccine manufacturing process, magnifection, devoid of the above-mentioned shortcomings and allowing consistent and efficient expression in plants of whole immunoglobulins (Igs).
  
  
  RESULTS
  Full idiotype (Id)-containing IgG molecules of 20 lymphoma patients and 2 mouse lymphoma models were expressed at levels between 0.5 and 4.8 g/kg of leaf biomass. Protein A affinity capture purification yielded antigens of pharmaceutical purity. Several patient Igs produced in plants showed specific cross-reactivity with sera derived from the same patients immunized with hybridoma-produced Id vaccine. Mice vaccinated with plant- or hybridoma-produced Igs showed comparable protection levels in tumor challenge studies.
  
  
  CONCLUSIONS
  This manufacturing process is reliable and robust, the manufacturing time from biopsy to vaccine is <12 weeks and the expression and purification of antigens require only 2 weeks. The process is also broadly applicable for manufacturing monoclonal antibodies in plants, providing 50- to 1000-fold higher yields than alternative plant expression methods.
authors.affiliation: Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Bendandi
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Marillonnet
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Kandzia
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Thieme
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Nickstadt
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Herz
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Fröde
authors.affiliation: Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Inogés
authors.affiliation: Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Lòpez-Dìaz de Cerio
authors.affiliation: Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Soria
authors.affiliation: Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
authors.forename: H
authors.initials: H
authors.lastname: Villanueva
authors.affiliation: Bayer BioScience, Gent, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Vancanneyt
authors.affiliation: Matrix Bioresearch, Vacaville, CA, USA.
authors.forename: A
authors.initials: A
authors.lastname: McCormick
authors.affiliation: DT/Consulting Group.
authors.forename: D
authors.initials: D
authors.lastname: Tusé
authors.affiliation: Bayer Schering Pharma, Wuppertal, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Lenz
authors.affiliation: Bayer Innovation GmbH, Duesseldorf, Germany.
authors.forename: J-E
authors.initials: JE
authors.lastname: Butler-Ransohoff
authors.affiliation: Icon Genetics GmbH, Halle, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Klimyuk
authors.affiliation: Icon Genetics GmbH, Halle, Germany. Electronic address: gleba@icongenetics.de.
authors.forename: Y
authors.initials: Y
authors.lastname: Gleba
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000818:Animals; D019496:Cancer Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D004526:Efficiency; D018506:Gene Expression Regulation, Plant; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D007206:Individuality; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology* / Q000628:therapy*; D051379:Mice; D008809:Mice, Inbred C3H; D046549:Plantibodies / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D019496:Cancer Vaccines; D007130:Immunoglobulin Idiotypes; D046549:Plantibodies; D014614:Vaccines, Synthetic