Protein nanoparticles (NPs) can be used as vaccine platforms for target antigen presentation. Aim: To conduct a proof-of-concept study to demonstrate that an effective NP platform can be built based on a short self-assembling peptide (SAP) rather than a large self-assembling protein. Materials & methods: SUMO-based protein fusions (SFs) containing an N-terminal SAP and a C-terminal antigen were designed, expressed in Escherichia coli and purified. The structure was investigated by electron microscopy. The antibody response was tested in mice after two adjuvant-free immunizations. Results: Renatured SFs form fiber-like NPs with the antigen exposed on the surface and induce a significant antibody response with a remarkably high target-to-platform ratio. Conclusion: The platform is effective and has considerable potential for modification toward various applications, including vaccine development.
A strain of the yeast Komagataella kurtzmanii, a producer of recombinant peptidase SerP38 from the yellow mealworm Tenebrio molitor, has been obtained. The level of proenzyme secretion was 20-50 mg/L. It was shown that the target His 6 -tagged protein was produced in two forms during secretion in yeast. One of them was a monomer that was efficiently purified via Ni-NTA chromatography and then activated with trypsin. Another form accumulated in the culture medium as oligomers prone to aggregation in the presence of Ni 2+ ions and was not activated by trypsin treatment. Aggregation is likely the result of the polypeptide-chain misfolding.
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