| Abstract Detail
Plant biotech & Risk Assessment Lai, Huafang [1], Khalsa, Guruatma [1], Arntzen, Charles [1], Chen, Qiang [2]. Development of a scalable extraction and purification strategy for MAbs and MAb-fusion proteins from transgenic Nicotiana benthamiana. Over the last two decades, plant-produced monoclonal antibodies (MAbs) have become a mature technology. Researchers have now produced over 100 different antibodies of IgG, IgA, dimeric IgA and SIgA isotype from many plant species. To date, all MAbs have shown antigen-binding activity and potency equivalent or similar to mammalian produced MAbs. In spite of this progress, remaining barriers still prevent the broad application of this technology platform. The lack of efficient and scalable extraction and purification strategy is one of these obstacles. Even though Protein A chromatography provides an excellent step for MAb purification, direct application of clarified Nicotiana benthamiana or Nicotiana tobacum extract to Protein A resin results in low MAb binding and recovery. In this study, we investigated the effect of a variety of extraction, filtration and chromatography steps on the removal of Protein-A-binding-interfering molecules from N. benthamiana extract. Our results indicate that tangential flow filtration and Ion-exchange chromatography are effective methods to remove the interfering molecules, thereby, increase the recovery of MAb and MAb-fusion protein from Protein A chromatography. The scalability and robustness of these methods suggest their applications to other plant-derived therapeutics.
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1 - Arizona State University, Center for Infectious Diseases and Vaccinology, Biodesign Institute
2 - Arizona State University, Center for Infectious Diseases and Vaccinology, Biodesign Institute, Department of Applied Biological Sciences, 1001 S. McAllister Avenue, Mail Zone 5401, Tempe, Arizona, 85287-5401, USA
Keywords:
MAbs
purification
Plant-derived therapeutics.
Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P45015
Abstract ID:1092 |
