Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants
Apart from tradition, the biospecific affinity isolation has become one of the most rapidly growing concerns with cellulose binding domains (CBDs), a high-capacity tag for cost-effective purifications of fusion proteins. We report here a new strategy optimized for isolation of two fusion proteins, the FGF-1 (a human functional protein) and the H5N1 (a vaccine candidate antigen) tagged with CBD were grown in transient Nicotiana benthamiana and transgenic Arabidopsis thaliana respectively. A notable fraction of the recombinant proteins was lost through plant debris pelleted from the plant-slurry made. However, this issue was resolved by adjusting tissue-to-buffer ratios with 1:10 and 1:15 in those plants respectively. Washing efficiencies were improved by agitating column beds with acidic buffer (20mM NaAc. pH 4.0) in Nicotiana and alkaline buffer (10mM Tris-base pH 8.0) in Arabidopsis. Adsorption and coupling of tagged proteins on cellulose matrices were affected by the buffer-logged resins. The column-beds, after pumping the moisture out, showed efficient in binding of antigens with almost no losses detected by immunoblot signals. The bound antigens were released efficiently from the cellulose matrices by 1% Cellobiose and 2% Triethylamine respectively. The successive purifications of these antigenic proteins with identical tags likely indicate the efficiency of the proposed strategy in providing a generic and cost-effective method to purify fusion proteins propagated in transgenic plants.
HMM Tariq Hossain,
Kwan Yong Choi,
Cellulose-Affinity Isolation of Vaccine Candidate Antigens from Transgenic Plants, Chemical and Biomolecular Engineering.
Vol. 3, No. 1,
2018, pp. 1-10.
Chan H. T., Daniell, H., 2015. Plant-made oral vaccines against infectious diseases – Are we there yet? Pl. Biotech. J.13: 1056-1070.
Korban S. S., 2002. Targeting and expression of antigenic proteins in transgenic plants for production of edible vaccines. In vitro Cellular and Developmental Biol.– Plant 38: 231-236.
Wycoff K. L., 2005. Secretory IgA antibodies from plants. Current Pharmaceutical Design 11: 2429-2437.
Fischer R., Schillberg S. (Eds.) 2004. Molecular Farming: Plant-made Pharmaceuticals and Technical Proteins WILLEY-VCH Verlog GmbH & Co., KGaA, Weinheim.
Streatfield J., Howard J. A., 2003. Plant Production Systems for Vaccines. Expert Review of Vaccines 2: 763-775.
Fiske M. J., Fredenburg R. A., VabDerMeid K. R., McMichael J. C., Arumugham R, 2001. Method for Reducing Endotoxin in Moraxella catarrhalis UspA2 Protein Preparations, J. Chromatogr. B Biomed. Sci Appl.753:269-278.
Magalhaes P. O., Lopes A. M., Mazzola P. G., Rangel-Yuagui C., Penna T. C., Pessoa A. Jr., 2007. Methods of Endotoxin Removal from Biological Preparations, a review, J. Pharm. Pharm. Sci.10: 388-404.
Hirayama C., Sakata M., 2002. Chromatographic removal of endotoxin from protein solutions by polymer particles. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.781: 419-432.
Herbes K., Sonnewald U., 1999. Production of new / modified proteins in transgenic plants. Current Opin. in Biotech. 10: 163-168.
Conard U., Fiedler U., 1998. Compartment specific accumulation of recombinant immunoglobulins in plant cells: an essential tool for antibody production and immunomodulation of physiological functions and pathogen activity. Pl. Mol. Biol. 38: 101-109.
Arntzen C. J., 1998. Pharmaceutical foodstuffs-oral immunization with transgenic plants. Nat. Med. Vaccine Suppl. 4: 502-503.
Schillberg S., Emans N., Fischer R., 2002. Antibody molecular farming in plants and plant cells. Phytochem. Rev.1: 45-54.
Stoger, E., Sack, M., Fischer, R., Christou, P., 2002. Plantibodies: applications, advantages and bottlenecks. Curr. Opin. Biotechnol.13(2):161-166.
Stoger, E., Sack, M., Nicholson L., Fischer, R., Christou, P., 2005. Recent Progress in Plantibody Technology. Current Pharma. Design11: 2439-2457.
Hellwigs S., Drossard J. Twyman R. M., Fischer R., 2004. Plant cell cultures for the production of recombinant proteins. Nature Biotech. 22(11): 1415-1422.
Smith D. B., Johnson, K. S., 1998. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67: 31-40.
Hedouelle, H., Duplay, P., 1998. Production in Escherichia coli and one-step purification of bifunctional hybrid proteins which bind maltose. Export of the Klenow polymerase into the periplasmic space. Eur. J. Biochem.17: 541-549.
Porath, J. Carlsson, J., Olsson, I., Belfrage, G., 1975. Metal Chelate affinity Chromatography, a new approach to protein purification. Nature 258: 598-599.
Lichty, J. J., Malecki, J. L., Agnew, H. D., Michelson-Horowitz, D. J., Tan, S., 2005. Comparision of affinity tags for protein purification. Protein Express. Purif. 41: 98-105.
Shani, Z., Shoseyov, O., 2001. Process of expressing and isolating recombinant proteins and recombinant protein products from plants, plant derived tissues or cultured cells. US patent 6: 331- 416.
Shoseyov, O., Shani, Z., Levy, I., 2006. Carbohydrate binding modules: biochemical properties and novel applications. Microbiol. Mol. Biol. Rev. 70(2): 283-95.
Rodriguez, B., Kavoosi, M., Koska, J., Creagh, A. L., Kilburn, D. G., Haynes, C. A. 2004. Inexpensive and generic affinity purification of recombinant proteins using a family 2a CBM fusion tag. Biotechnol. Prog. 20(5): 1479-89.
Hearn, M. T. W., Acosta, D., 2001. Applications of novel affinity cassette methods: Use of peptide fusion handles for the purification of recombinant proteins. J. Mol. Recognit. 14: 323-369.
Evangelista, R. L., Kusnadai, A. R., Howard, J. A., Nikolov, Z. L., 1998. Process and economic evaluation of the extraction and purification of recombinant beta-glucoronidase from transgenic corn. Biotechnol Prog. 14: 607-614.
Stoger, E., Sack, M., Nicholson, L. Fischer, R., Christou, P., 2005. Recent progress in plantibody technology. Curr. Pharm. Design 11: 2439-2457.
Witcher, D., Hood, E. E., Peterson, D., Baily, M., Bond, D., Kusandai, A., Evangelista, R., Nikolov, Z., Wooge, C., Mehigh, R., Kappel, W. Register, J. C., Howard, J. A., 1998. Commercial production of β-glucoronidase (GUS): a model system for the production of protein in plants. Mol. Breed. 4: 301-312.
Conard, U., Fiedler, U. 1998. Compartment-specific accumulation of recombinant immunoglobulins in plant cells: as essential tool for antibody production and immunomodulation of physiological functions and pathogen activity Pl. Mol. Biol. 38: 101-109.
Voinnet O, Rivas S, Mestre P, Baulcombe D., 2003. An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Pl. J. 33: 949-956.
Platis, D., Drossard, J., Fischer, R., Ma, J. K.-C., Labrou, N. E., 2008. New downstream processing strategy of the purification of monoclonal antibodies from transgenic tobacco plants. J. Chromatogr. A: 80-89.
Fischer, R., Emans, N., 2000. Molecular farming of pharmaceutical proteins. Transgenic Res. 9: 279-299.
Wilken, L. R., Nikolov, Z. L., 2012. Recovery and purification of plant-made recombinant proteins. Biotechnol. Adv. 30: 419-433.
Gottschalk, U., 2008. Bioseparation in antibody manufacturing: the good: the bad and the ugly. Biotechnol. Progr. 24: 496-503.
Nicolov, Z. L. Regan, J. R., Dcikey, L. F., Woodard, S. L., 2009. Purification of antibodies from transgenic plants. In: Gottschalk U., editor. Process scale purification of antibodiesWiley-VCH:387-406.
Barros, G. O. F., Woodard, S. L., Nikolov, Z. L., 2011. Phenolics removal from transgenic Lemma minor extracts expressing mAb and impact on mAb production cost. Biotechnol. Progr. 27: 410-418.
Nykiforuk, C. L., Shen, Y., Murray, E. W., Boothe, J. G., Bussenil, D. et al., 2011. Expression and recovery of biologically active recombinant Apolipoprotein Al Milano from transgenic safflower (C. tinctorius) seeds. Pl. Biotechnol. 9: 250-263.
Holler, C., Zhang, C., 2007. Purification of an acidic recombinant protein from transgenic tobacco. Biotechnol. Bioeng. 99(4): 902-909.
Hong, J., Ye, X., Wang, Y., Zhang, Y.-H. P., 2008. Bioseparation of recombinant cellulose-binding module-proteins by affinity adsorption on an ultra-high-capacity cellulose adsorbent. Analytica Chemica Acta 621: 193-199.
Varela, M. Kimmel, P. L., Phillips, T. M., Mishkin, G. J. Lew, S. Q., Bosch, J. P., 2001. Biocompatibility of hemodialysis membranes: interrelations between plasma complement and cytokine levels. Blood Purif. 19:370-379.
Hong, J., Wang, Y., Ye, X., Zhang, Y.-H. P., 2008. Simple protein puriﬁcation through afﬁnity adsorption on regenerated amorphous cellulose followed by intein self-cleavage. J. Chromatogr. A: 150-154.
Shoseyov, O., Shani, Z., Levy, I., 2006. Carbohydrate Binding Modules: Biochemical Properties and Novel Applications. Microbiol. Mol. Biol. Rev. 70 (2):283- 295.