Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A
Open Access
- 21 November 2006
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 103 (47), 17678-17683
- https://doi.org/10.1073/pnas.0608869103
Abstract
Earlier attempts to express peptides longer than 20 aa on the surface of tobamoviruses such as tobacco mosaic virus have failed. Surprisingly, we found that a functional fragment of protein A (133 aa) can be displayed on the surface of a tobamovirus as a C-terminal fusion to the coat protein via a 15-aa linker. The macromolecular nature of these nanoparticles allowed the design of a simple protocol for purification of mAbs with a recovery yield of 50% and >90% product purity. The extremely dense packing of protein A on the nanoparticles (>2,100 copies per viral particle) results in an immunoadsorbent with a binding capacity of 2 g mAb per g. This characteristic, combined with the high level of expression of the nanoparticles (>3 g adsorbent per kg of leaf biomass), provides a very inexpensive self-assembling matrix that could meet the criteria for a single-use industrial immunoadsorbent for antibody purification.Keywords
This publication has 42 references indexed in Scilit:
- Mutational analysis of the major coat protein of M13 identifies residues that control protein displayProtein Science, 2008
- Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectorsProceedings of the National Academy of Sciences of the United States of America, 2006
- Plant biopharming of monoclonal antibodiesVirus Research, 2005
- Antibodies and Genetically Engineered Related Molecules: Production and PurificationBiotechnology Progress, 2004
- Tobacco Mosaic Virus: Pioneering Research for a CenturyTHE PLANT CELL ONLINE, 1999
- Completion of a cDNA sequence from a tobamovirus pathogenic to crucifersGene, 1995
- Systemic production of foreign peptides on the particle surface of tobacco mosaic virusFEBS Letters, 1995
- Complete nucleotide sequence and genome organization of a tobamovirus infecting cruciferae plantsFEBS Letters, 1994
- Production of enkephalin in tobacco protoplasts using tobacco mosaic virus RNA vectorFEBS Letters, 1990
- Visualization of protein-nucleic acid interactions in a virus: Refined structure of intact tobacco mosaic virus at 2.9 Å resolution by X-ray fiber diffractionJournal of Molecular Biology, 1989