Rapid, high-yield production in plants of individualized idiotype vaccines for non-Hodgkin's lymphoma.
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.
Bendandi
M
M
Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Marillonnet
S
S
Icon Genetics GmbH, Halle, Germany.
Kandzia
R
R
Icon Genetics GmbH, Halle, Germany.
Thieme
F
F
Icon Genetics GmbH, Halle, Germany.
Nickstadt
A
A
Icon Genetics GmbH, Halle, Germany.
Herz
S
S
Icon Genetics GmbH, Halle, Germany.
Fröde
R
R
Icon Genetics GmbH, Halle, Germany.
Inogés
S
S
Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Lòpez-Dìaz de Cerio
A
A
Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Soria
E
E
Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Villanueva
H
H
Laboratory of Immunotherapy, Oncology Division, Center for Applied Medical Research, University of Navarra, Pamplona, Spain.
Vancanneyt
G
G
Bayer BioScience, Gent, Belgium.
McCormick
A
A
Matrix Bioresearch, Vacaville, CA, USA.
Tusé
D
D
DT/Consulting Group.
Lenz
J
J
Bayer Schering Pharma, Wuppertal, Germany.
Butler-Ransohoff
J-E
JE
Bayer Innovation GmbH, Duesseldorf, Germany.
Klimyuk
V
V
Icon Genetics GmbH, Halle, Germany.
Gleba
Y
Y
Icon Genetics GmbH, Halle, Germany. Electronic address: gleba@icongenetics.de.
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
D019496:Cancer Vaccines; D007130:Immunoglobulin Idiotypes; D046549:Plantibodies; D014614:Vaccines, Synthetic