Immunoabsorbent nanoparticles based on a tobamovirus displaying protein A

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 Cterminal 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.antibody purification ͉ coat protein fusion ͉ viral particle

Control of peptide nanotube diameter by chemical modifications of an aromatic residue involved in a single close contact

“…There has been rapid progress in the development of nanomaterials such as nanowires, nanotubes, nanoparticles, and nanocrystals, which show superior electronic, magnetic, optical, and mechanical properties as compared with their bulk counterparts. These materials are extremely attractive as building blocks for various applications, as, for instance, electronic and magnetic devices (2), catalysis (3), nanolasers (4), nanofluidics (5), nanolithography (6) drug delivery, tissue engineering (7), biosensors (8)(9)(10), and medicine (11). A very interesting aspect of these nanomaterials is that their physical and chemical properties are tunable via the control of their size and shape (12).…”

Role of plant expression systems in antibody production for passive immunization

“…Also, Agrobacterium infiltration entails the presence of a large amount of bacterial cells, introducing the risk of bacterial endotoxin contamination. However, innovative, cost-effective purification systems, like the tobamovirus-protein A fusion, are being developed (Werner et al, 2006). Taken all together, the tobacco leaf-based expression system, both stable as well as transient, seems very promising.…”

Metabolic engineering of plants using a disarmed potyvirus vector

“…The triple module magnICON system was used to express several proteins in plants, with special attention on monoclonal antibodies (mAbs) Webster et al, 2009;Huang et al, 2006). Since TMV capsid is made-up from more than 2000 copies of the CP and both the N-and the C-terminus of the CP are situated on the virion's surface, magnICON was also successfully used as an epitope presentation system (Turpen et al, 1995;Wigdorovitz et al, 1999;Werner et al, 2006;Noris et al, 2011). Other important gene expression system based on TMV is TRBO which is comparable to magnICON in terms of recombinant protein yields, but simpler in design (Lindbo, 2007).…”