A research lately revealed in Communications Biology (Nature Portfolio) by Esra Oktay, Farhang Alem, Keziah Hernandez, Michael Girgis, Christopher Inexperienced, Divita Mathur, Igor L. Medintz, Aarthi Narayanan, and Rémí Veneziano introduces an revolutionary DNA origami–primarily based nanovaccine platform focusing on the receptor-binding area (RBD) of SARS-CoV-2. Their findings spotlight the potential of rationally engineered DNA nanoparticles to elicit sturdy and sturdy immune safety.
The researchers designed wireframe DNA origami nanoparticles (DNA-NPs) functionalized with ten trimerized RBD antigens and CpG oligonucleotides as adjuvants. Trimer meeting was achieved by peptide nucleic acid (PNA)–mediated conjugation of protein G to RBD-Fc, enabling exact orientation on DNA-NP overhangs. Biophysical characterization employed dynamic gentle scattering, Förster resonance vitality switch (FRET)–primarily based stability assays, and quantitative fluorescence labeling. Structural affirmation was obtained by way of atomic pressure microscopy (AFM). For prime-resolution AFM imaging, the staff employed NanoWorld® USC-F0.3-k0.3 ultra-short cantilevers, chosen for his or her 300 kHz resonance frequency, ~0.3 N/m pressure fixed, and <10 nm tip radius. These probes supplied enhanced stability and backbone, enabling correct nanoscale visualization of the vaccine constructs.
In murine fashions, the totally assembled RBD-DNA-NP with CpG supplied full safety following viral problem. Vaccinated mice exhibited no weight reduction or mortality, in stark distinction to regulate teams that acquired unconjugated parts, naked origami scaffolds, or RBD alone—a number of of which skilled important morbidity or mortality. Immunological assays revealed sustained neutralizing antibody exercise two months post-immunization, with sturdy titers of RBD-specific IgG, IgM, and IgA, underscoring the sturdiness of the immune response.
This work demonstrates that DNA origami nanostructures can function an adaptable and efficient vaccine supply platform. By integrating exact antigen show with adjuvant loading, the strategy gives a promising blueprint for next-generation nanovaccines. The usage of superior AFM probes, such because the NanoWorld® USC-F0.3-k0.3, additional underscores the significance of high-precision nanometrology instruments in validating nanoscale biomedical designs.
a Hydrodynamic diameter measurement of DNA-NPs with DLS. b Atomic pressure microscopy imaging of DNA-NPs formation and conjugation with 3-mer RBDs (scale bar: 20 nm). c Fluorescence-intensity primarily based dedication of PG and RBD stoichiometry on DNA origami NP. i) Tryptophan fluorescence emission was used to find out the full variety of PG loaded on the floor of the DNA-NPs. ii) Second, the stoichiometry of RBD on NP was quantified by way of measuring the emission of Cy5 dyes conjugated to the RBD antigens. The bar graph represents the full protection proportion for the PG (inexperienced bar) and the 3-mer RBD (orange bar) on the DNA-NP floor normalized to the variety of conjugation websites obtainable. Knowledge are proven because the imply ± SD (n = 3 unbiased experiments).
This text comprises photographs reused from Jia et al., Nature Communications 14, 1394 (2023), licensed beneath CC BY 4.0.