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Purification of the therapeutic antibody trastuzumab from genetically modified plants using safflower Protein A-oleosin oilbody technology
- Source :
- Transgenic Research. 21:1291-1301
- Publication Year :
- 2012
- Publisher :
- Springer Science and Business Media LLC, 2012.
-
Abstract
- Production of therapeutic monoclonal antibodies using genetically modified plants may provide low cost, high scalability and product safety; however, antibody purification from plants presents a challenge due to the large quantities of biomass that need to be processed. Protein A column chromatography is widely used in the pharmaceutical industry for antibody purification, but its application is limited by cost, scalability and column fouling problems when purifying plant-derived antibodies. Protein A-oleosin oilbodies (Protein A-OB), expressed in transgenic safflower seeds, are relatively inexpensive to produce and provide a new approach for the capture of monoclonal antibodies from plants. When Protein A-OB is mixed with crude extracts from plants engineered to express therapeutic antibodies, the Protein A-OB captures the antibody in the oilbody phase while impurities remain in the aqueous phase. This is followed by repeated partitioning of oilbody phase against an aqueous phase via centrifugation to remove impurities before purified antibody is eluted from the oilbodies. We have developed this purification process to recover trastuzumab, an anti-HER2 monoclonal antibody used for therapy against specific breast-cancers that over express HER2 (human epidermal growth factor receptor 2), from transiently infected Nicotiana benthamiana. Protein A-OB overcomes the fouling problem associated with traditional Protein A chromatography, allowing for the development of an inexpensive, scalable and novel high-resolution method for the capture of antibodies based on simple mixing and phase separation.
- Subjects :
- medicine.drug_class
Transgene
Molecular Sequence Data
Carthamus tinctorius
Nicotiana benthamiana
Antibodies, Monoclonal, Humanized
Monoclonal antibody
Chromatography, Affinity
Plantibodies
Column chromatography
Tobacco
Genetics
medicine
Humans
Amino Acid Sequence
Staphylococcal Protein A
Organelles
biology
Arabidopsis Proteins
Trastuzumab
Plants, Genetically Modified
biology.organism_classification
Fusion protein
Molecular biology
Biochemistry
biology.protein
Animal Science and Zoology
Oleosin
Antibody
Protein A
Agronomy and Crop Science
Biotechnology
Subjects
Details
- ISSN :
- 15739368 and 09628819
- Volume :
- 21
- Database :
- OpenAIRE
- Journal :
- Transgenic Research
- Accession number :
- edsair.doi.dedup.....74d2101e8d7d9c8a18a726ab6ecee7c6