ITQB: ECAL- Master Theses
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- Tyrosinase-catalysed bioconjugation for the production of ferritin - based vaccine candidatesPublication . Patão, Sara Rae Jones; Roldão, António"Ferritin has recently gathered attention as a vaccine platform due to desirable thermal/chemical properties and presumable safety owing to its biocompatibility. However, the expression of antigens fused to protein carriers is correlated with structural destabilisation. By operating under mild conditions with fast kinetics and high selectivity, enzymatic-mediated linkage of biomolecules enables retention of folding and function while allowing for high modifcation yields. One example is the tyrosinase-catalysed bioconjugation strategy, where tyrosine-tagged peptides are linked to cysteine sidechains at the surface of a protein scaffold. This work describes for the frst time the use of Agaricus bisporus tyrosinase (AbTYR) to bind tyrosine-tagged SARS-CoV-2 receptor-binding domain (RBD-Y), produced in insect cells, to native cysteines at the surface of human ferritin (HFt) nanoparticles produced in E. Coli. An initial assessment of tyrosinase-mediated bioconjugation of ferritin to RBD-Y revealed effciencies of ~68 %, encouraging the design of optimisation strategies.(...)"