The self-assembling nature of coiled-coils has brought this common structural motif into the spotlight of protein design since it offers a customizable framework for engineering innovative protein nanoparticles with tailored functionalities. We recently harnessed the self-assembling capabilities of ZapB, a bacterial coiled-coil protein, to build up fluorescent protein nanoparticles possessing remarkable affinity for antibodies. Here, we describe a complete workflow detailing the design, production, and characterization of such coiled-coil-based protein nanostructures. Additionally, we detail their functionalization with specific antibodies and illustrate their utility in stimulating the activation and proliferation of human T cells, underscoring the potential of these protein-only nanoparticles in immunotherapeutic interventions.
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