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 progressive DNA origami–primarily based nanovaccine platform concentrating 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 mild scattering, Förster resonance power switch (FRET)–primarily based stability assays, and quantitative fluorescence labeling. Structural affirmation was obtained through atomic power microscopy (AFM). For top-resolution AFM imaging, the group employed NanoWorld® USC-F0.3-k0.3 ultra-short cantilevers, chosen for his or her 300 kHz resonance frequency, ~0.3 N/m power fixed, and <10 nm tip radius. These probes offered enhanced stability and backbone, enabling correct nanoscale visualization of the vaccine constructs.
In murine fashions, the totally assembled RBD-DNA-NP with CpG offered 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 vital 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 method offers a promising blueprint for next-generation nanovaccines. Using 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 power 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 through measuring the emission of Cy5 dyes conjugated to the RBD antigens. The bar graph represents the full protection share 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 incorporates photos reused from Jia et al., Nature Communications 14, 1394 (2023), licensed underneath CC BY 4.0.
