Assembly and Assessment of DNA Scaffolded Vaccines
Vaccines play an important role in preventing many life-threatening infectious diseases. To meet the demand of vaccination for treating a wide range of diseases, rational vaccine design has been recognized as a desirable and necessary strategy for develop
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Introduction Vaccination is one of the most cost-effective public health interventions. Due to its success in the conquest of many infectious diseases, vaccination has also been explored as a prevention and/or therapeutic strategy in dealing with many other diseases, including autoimmunity [1], cancer [2], and drug abuse [3]. Over the past three decades, recombinant DNA technology has significantly advanced the vaccine field, leading to safer recombinant microbes, DNA vaccines, and subunit vaccines. However, recombinant subunit vaccines usually lack sufficient efficacy. In recent years, the advancement in nanotechnology and the availability of various nanomaterials has made strides in improving the efficacy of the subunit vaccines [4]. Inspired by nature, synthetic microparticles and nanoparticles have been engineered to incorporate well-defined antigenic components and adjuvant molecules to form nanovaccines that can be rationally designed and tailored for enhanced immunogenicity and desired safety [4, 5]. Recently, DNA nanostructures have been recognized as an ideal structural material for the assembly of various biomolecules [6–12], including vaccines [13]. DNA nanotechnology makes good use of the simple Watson– Crick base pairing principle to provide a highly programmable and robust way to self-assemble diverse nanostructures [14]. Various two- and three-dimensional DNA nanostructures have been constructed [15–18], thereby providing a diverse “tool box,” and have been utilized for precise organization of biochemical molecules and targeted cellular transport [7, 10, 11]. DNA nanoscaffold provides control over structural features such as particle size and geometry, epitope valency and configuration, and has been recently explored as a synthetic platform for vaccine assembly, as well as assembly of other immunomodulating modules [8, 9, 13]. Here
Sunil Thomas (ed.), Vaccine Design: Methods and Protocols, Volume 2: Vaccines for Veterinary Diseases, Methods in Molecular Biology, vol. 1404, DOI 10.1007/978-1-4939-3389-1_21, © Springer Science+Business Media New York 2016
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Xiaowei Liu et al.
we describe the assembly of DNA scaffolded vaccines, the structural stability of these vaccines, and the assessment of their immunogenicity.
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Materials
2.1 Buffer Preparation
1. Stock (50× TAE) buffer: Mix the following components and add diH2O (distilled and deionized water) to a total volume of 1000 mL: 242.2 g Tris base (Formula weight [FW] 121.1, final concentration 2 M); 57.2 mL acetic acid (FW 60.05, final concentration 1 M); 37.2 g ethylenediaminetetraacetic acid disodium salt dihydrate (EDTA·Na2, FW 372.24, final concentration 0.1 M). Store at 4 °C. 2. Annealing (10× TAE/Mg2+) buffer: Weigh 26.8 g magnesium acetate tetrahydrate (FW 214.46, final concentration 125 mM), and mix with 200 mL stock (50× TAE) buffer. Add diH2O to a final volume of 1000 mL. Adjust pH to 8.0, and filter through bottle top vacuum filter (500 mL, pore size 0.22 μm). Store the annealing buffer in 1 L sterile plastic bottles or as
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