High-yield expression of recombinant soybean agglutinin in plants using transient and stable systems
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ORIGINAL PAPER
High-yield expression of recombinant soybean agglutinin in plants using transient and stable systems Reynald Tremblay • Mary Feng • Rima Menassa • Norman P. A. Huner Anthony M. Jevnikar • Shengwu Ma
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Received: 7 January 2010 / Accepted: 8 June 2010 / Published online: 18 June 2010 Ó Springer Science+Business Media B.V. 2010
Abstract Soybean agglutinin (SBA) is a specific N-acetylgalactosamine-binding plant lectin that can agglutinate a wide variety of cells. SBA has great potential for medical and biotechnology-focused applications, including screening and treatment of breast cancer, isolation of fetal cells from maternal blood for genetic screening, the possibility as a carrier system for oral drug delivery, and utilization as an affinity tag for high-quality purification of tagged proteins. The success of these applications, to a large degree, critically
Electronic supplementary material The online version of this article (doi:10.1007/s11248-010-9419-0) contains supplementary material, which is available to authorized users. R. Tremblay N. P. A. Huner S. Ma (&) Department of Biology, University of Western Ontario, London, ON N6A 5B7, Canada e-mail: [email protected] M. Feng Department of Physiology and Pharmacology, University of Western Ontario, London, ON N6A 5C1, Canada R. Menassa Agriculture and Agri-Food Canada, 1391 Sandford St., London, ON N5V 4T3, Canada
depends on the development of a highly efficient expression system for a source of recombinant SBA (rSBA). Here, we demonstrate the utility of transient and stable expression systems in Nicotiana benthamiana and potato, respectively, for the production of rSBA, with the transgenic protein accumulated to 4% of total soluble protein (TSP) in Nicotiana benthamiana leaves and 0.3% of TSP in potato tubers. Furthermore, we show that both plant-derived rSBAs retain their ability to induce the agglutination of red blood cells, are similarly glycosylated when compared with native SBA, retained their binding specificity for N-acetylgalactosamine, and were highly resistant to degradation in simulated gastric and intestinal fluids. Affinity column purification using N-acetylgalactosamine as a specific ligand resulted in high recovery and purity of rSBA. This work is the first step toward use of rSBA for various new applications, including the development of rSBA as a novel affinity tag for simplified purification of tagged proteins and as a new carrier molecule for delivery of oral drugs. Keywords Soybean agglutinin Nicotiana benthamiana Solanum tuberosum Hemagglutination N-Acetylgalactosamine Protein purification
A. M. Jevnikar S. Ma Transplantation Immunology Group, Lawson Health Research Institute, London, ON N6A 4G5, Canada
Introduction
S. Ma Plantigen Inc., 375 South Street, London, ON N6A 4G5, Canada
Soybean agglutinin (SBA) is a legume lectin glycoprotein that binds non-covalently to specific
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cell-surface carbohydrates, provoking agglutination of the bound cells when in solution. SBA has been used to fractionate dif
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