Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectors.
Plant viral vectors allow expression of heterologous proteins at high yields, but so far, they have been unable to express heterooligomeric proteins efficiently. We describe here a rapid and indefinitely scalable process for high-level expression of functional full-size mAbs of the IgG class in plants. The process relies on synchronous coinfection and coreplication of two viral vectors, each expressing a separate antibody chain. The two vectors are derived from two different plant viruses that were found to be noncompeting. Unlike vectors derived from the same virus, noncompeting vectors effectively coexpress the heavy and light chains in the same cell throughout the plant body, resulting in yields of up to 0.5 g of assembled mAbs per kg of fresh-leaf biomass. This technology allows production of gram quantities of mAbs for research purposes in just several days, and the same protocol can be used on an industrial scale in situations requiring rapid response, such as pandemic or terrorism events.
Giritch
Anatoli
A
Icon Genetics GmbH, Weinbergweg 22, 06120 Halle, Germany.
Marillonnet
Sylvestre
S
Engler
Carola
C
van Eldik
Gerben
G
Botterman
Johan
J
Klimyuk
Victor
V
Gleba
Yuri
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity / Q000276:immunology; D000941:Antigens / Q000276:immunology; D016466:CHO Cells; D002465:Cell Movement; D020090:Chromosome Segregation / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D018515:Plant Leaves / Q000166:cytology / Q000821:virology; D046549:Plantibodies / Q000378:metabolism*; D017863:Potexvirus / Q000502:physiology*; D012093:Replicon / Q000235:genetics; D014026:Nicotiana / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology; D000097573:Red Fluorescent Protein
D000911:Antibodies, Monoclonal; D000941:Antigens; D049452:Green Fluorescent Proteins; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D008164:Luminescent Proteins; D046549:Plantibodies; D000097573:Red Fluorescent Protein