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Viruses provide a whole new set of building blocks for the development of new materials and as such have found application in, for example, materials science, nanotechnology, and medicine. Their monodispersity and high degree of symmetry surpass any synthetic nanoparticle currently available, and due to the huge amount of different viruses found on Earth there are many shapes and sizes to choose from. In particular the capsid of viruses, that surrounds their genome, is commonly studied. This protein shell transports and protects the genome, and releases the viral genetic material once it has entered a host cell to allow reproduction of the viral components. After reproduction, these reassemble into full viruses which are released from the host. This assembly and disassembly behavior requires specific inter-subunit interactions to provide stability of the virus during transportation while allowing disassembly to infect a host. Both for the purpose of understanding viral reproduction and treating viral infections as well as for using viral components for various applications and being able to predict the structures that will be formed from them, gaining insight in these interactions is crucial. The work described in this thesis extends our knowledge on viral assembly, by studying the assembly and confinement conditions of the cowpea chlorotic mottle virus (CCMV). By introducing microscale thermophoresis (MST) as a new way to study (self-)assembly, we were able to study and compare the assembly behavior of native CCMV capsid protein (CP) with that of two genetically modified versions of this protein over a wide range of conditions (Chapter 3). MST was also used in combination with isothermal titration calorimetry (ITC) to study CCMV CP assembly into virus-like particles (VLPs) templated by polyanionic species at neutral pH (Chapter 4). Continuing the study of polyanion-templated assembly, we determined the minimum length of single-stranded (ss)DNA, correlating to a minimal electrostatic interaction, required to induce viral assembly at neutral pH (Chapter 5).

The Assembly and Confinement Properties of the Cowpea Chlorotic Mottle Virus