Abstract
Fmoc solid-phase peptide synthesis (SPPS) is the most common approach used to synthesize natural and unnatural peptides. However, the synthesis of sequences longer than 30–60 amino acids is associated with a drastic reduction in peptide quality. Thus, large and complex peptides are normally synthesized as fragments, which are then conjugated together. Here, we describe the synthesis of a large, branched peptide, a multi-epitope vaccine candidate against Group A Streptococcus, with the help of microwave-assisted Fmoc-SPPS, thiol-maleimide conjugation, and copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) “click” reaction.
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References
Skwarczynski M, Toth I (2016) Peptide-based synthetic vaccines. Chem Sci 7(2):842–854. https://doi.org/10.1039/c5sc03892h
Massell BF (1969) Rheumatic fever following streptococcal vaccination. JAMA 207(6). https://doi.org/10.1001/jama.1969.03150190037007
Marijon E, Mirabel M, Celermajer DS, Jouven X (2012) Rheumatic heart disease. Lancet 379(9819):953–964. https://doi.org/10.1016/s0140-6736(11)61171-9
Rajčáni J, Szathmary S (2018) Peptide vaccines: new trends for avoiding the autoimmune response. Open Infect Dis J 10(1):47–62. https://doi.org/10.2174/1874279301810010047
Li W, Joshi MD, Singhania S, Ramsey KH, Murthy AK (2014) Peptide vaccine: progress and challenges. Vaccines (Basel) 2(3):515–536. https://doi.org/10.3390/vaccines2030515
Skwarczynski M, Toth I (2011) Lipid-core-peptide system for self-adjuvanting synthetic vaccine delivery. Methods Mol Biol 751:297–308. https://doi.org/10.1007/978-1-61779-151-2_18
Zhong W, Skwarczynski M, Toth I (2009) Lipid core peptide system for gene, drug, and vaccine delivery. Aust J Chem 62(9):956–967. https://doi.org/10.1071/ch09149
Moyle PM, Toth I (2013) Modern subunit vaccines: development, components, and research opportunities. ChemMedChem 8(3):360–376. https://doi.org/10.1002/cmdc.201200487
Alexander J, Sidney J, Southwood S, Ruppert J, Oseroff C, Maewal A, Snoke K, Serra HM, Kubo RT, Sette A, Grey HM (1994) Development of high potency universal DR-restricted helper epitopes by modification of high affinity DR-blocking peptides. Immunity 1(9):751–761. https://doi.org/10.1016/s1074-7613(94)80017-0
Erskine CL, Krco CJ, Hedin KE, Borson ND, Kalli KR, Behrens MD, Heman-Ackah SM, von Hofe E, Wettstein PJ, Mohamadzadeh M, Knutson KL (2011) MHC class II epitope nesting modulates dendritic cell function and improves generation of antigen-specific CD4 helper T cells. J Immunol 187(1):316–324. https://doi.org/10.4049/jimmunol.1100658
Moyle PM, Hartas J, Henningham A, Batzloff MR, Good MF, Toth I (2013) An efficient, chemically-defined semisynthetic lipid-adjuvanted nanoparticulate vaccine development system. Nanomedicine 9(7):935–944. https://doi.org/10.1016/j.nano.2013.01.009
Ahmad Fuaad AA, Skwarczynski M, Toth I (2016) The use of microwave-assisted solid-phase peptide synthesis and click chemistry for the synthesis of vaccine candidates against hookworm infection. Methods Mol Biol 1403:639–653. https://doi.org/10.1007/978-1-4939-3387-7_36
Celebioglu A, Uyar T (2011) Electrospun porous cellulose acetate fibers from volatile solvent mixture. Mater Lett 65(14):2291–2294. https://doi.org/10.1016/j.matlet.2011.04.039
Dyke JM, Groves AP, Morris A, Ogden JS, Dias AA, Oliveira AMS, Costa ML, Barros MT, Cabral MH, Moutinho AMC (1997) Study of the thermal decomposition of 2-azidoacetic acid by photoelectron and matrix isolation infrared spectroscopy. J Am Chem Soc 119(29):6883–6887. https://doi.org/10.1021/ja964354v
Bretherick L (1990) Specific chemicals. In: Bretherick L (ed) Bretherick’s handbook of reactive chemical hazards. Butterworth-Heinemann, Oxford, pp 1–1475. https://doi.org/10.1016/b978-0-7506-0706-3.50009-2
Wehrstedt KD, Wandrey PA, Heitkamp D (2005) Explosive properties of 1-hydroxybenzotriazoles. J Hazard Mater 126(1–3):1–7. https://doi.org/10.1016/j.jhazmat.2005.05.044
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Dai, C., Stephenson, R.J., Skwarczynski, M., Toth, I. (2020). Application of Fmoc-SPPS, Thiol-Maleimide Conjugation, and Copper(I)-Catalyzed Alkyne-Azide Cycloaddition “Click” Reaction in the Synthesis of a Complex Peptide-Based Vaccine Candidate Against Group A Streptococcus. In: Hussein, W., Skwarczynski, M., Toth, I. (eds) Peptide Synthesis. Methods in Molecular Biology, vol 2103. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0227-0_2
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DOI: https://doi.org/10.1007/978-1-0716-0227-0_2
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