In Situ Covalent Functionalization of DNA Origami Virus-like Particles

Abstract

DNA origami is a powerful nanomaterial for biomedical applications due in part to its capacity for programmable, site-specific functionalization. To realize these applications, scalable and efficient conjugation protocols are needed for diverse moieties ranging from small molecules to biomacromolecules. Currently, there are no facile and general methods for in situ covalent modification and label-free quantification of reaction conversion. Here, we investigate the postassembly functionalization of DNA origami and the subsequent high-performance liquid chromatography-based characterization of these nanomaterials. Following this approach, we developed a versatile DNA origami functionalization and characterization platform. We observed quantitative in situ conversion using widely accessible click chemistry for carbohydrates, small molecules, peptides, polymers, and proteins. This platform should provide broader access to covalently functionalized DNA origami, as illustrated here by PEGylation for passivation and HIV antigen decoration to construct virus-like particle vaccines.

Anion-specific structure and stability of guanidinium-bound DNA origami

“…Moreover, despite having been studied for several decades, even the ubiquitously employed Gdm + -induced denaturation of proteins is not completely understood yet [24] , [25] , [26] , [27] . From an application-oriented point of view, the interaction of chemical denaturants with DNA is highly relevant for various processes such as isothermal and low-temperature DNA origami assembly [28] , [29] , assembly of DNA origami nanostructures from double-stranded (ds) DNA [30] and intact bacteriophages [31] , selective DNA origami denaturation for analytical purposes [32] , and the removal of DNA origami masks in molecular lithography [33] , [34] . In this context, Gdm + is particularly interesting because its effect on DNA origami nanostructures is strongly influenced by concentration, temperature, and the presence and concentration of other ions [19] , [23] , which may be exploited for fine-tuning its activity to precisely match the requirements of a given application.…”

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

“…We hypothesized that such nanoscale organization could promote TD antibody responses against protein antigens but confine scaffold-directed B-cell responses to the non-boostable, extrafollicular pathway devoid of immunological memory. Wireframe DNA origami provides access to designer VLPs of controlled geometry and size at the 20 to 200 nm scale with independently programmable geometry, valency and stoichiometry of antigen display [42][43][44][45][46] . We and others recently leveraged this platform to probe the nanoscale parameters of IgM recognition and of BCR signaling in reporter B-cell lines, suggesting that increased antigen spacing up to 30 nm promotes early B-cell activation [47][48] .…”

Aptamers as promising nanotheranostic tools in the COVID‐19 pandemic era

“…In addition, low aptamer threshold would be required to prevent or reduce the virus infection in the lower and upper respiratory airways. To achieve this, medically relevant customizable molecular scaffolds (Knappe et al, 2021 ; Kwon et al, 2020 ; Veneziano et al, 2020 ) and biocompatible particulate nanotechnology platforms (Lauster et al, 2017 , 2020 ; Ogata et al, 2016 ; Ueda et al, 2020 ) with tremendous promises can be utilized to incorporate both sensing, inhibiting and neutralizing aptamers to address above‐mentioned challenges on both prophylactics and treatment fronts (Figure 5a ). These include liposomes, viral‐like particles, polymeric nanoparticles/micelles, dendrimers, protein cages, lipid nanoparticles, quantum dots, and DNA nanostructures.…”