Characterization of Hybrid Virus-Like Particles

The VLPs were characterized as previously described^{27 (link),28 (link),29 (link)}. The VLP concentration was determined using a Pierce BCA assay kit (Thermo Fisher Scientific). To confirm hybrid QβS100A9 assembly and peptide display, 10 μg of QβS100A9 particles was analyzed by SDS-PAGE under reducing conditions on NuPAGE 12% Bis-Tris protein gels (Thermo Fisher Scientific) stained with GelCode Blue Safe protein stain (Thermo Fisher Scientific). The gel images were acquired using the ProteinSimple FluorChem R imaging system, and densitometry was used to determine the number of peptides displayed per hybrid QβS100A9 VLP. The integrity of VLPs was confirmed by TEM using a FEI Tecnai Spirit G2 BioTWIN instrument to examine samples stained with 2% uranyl acetate. FPLC was carried out using an AKTA-FPLC 900 system fitted with Superose 6 Increase 10/300 GL columns (GE Healthcare) using PBS as the mobile phase. Particle size was confirmed by DLS using a Malvern Instruments Zetasizer Nano at 25 °C and plastic disposable cuvettes.

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Ortega-Rivera O.A., Shin M.D., Moreno-Gonzalez M.A., Pokorski J.K, & Steinmetz N.F. (2022). A single-dose Qβ VLP vaccine against S100A9 protein reduces atherosclerosis in a preclinical model. Advanced therapeutics, 5(10), 2200092.

Publication 2022

Pierce BCA assay kit NuPAGE 12% Bis-Tris protein gels GelCode Blue Safe protein stain Superose 6 Increase 10/300 GL columns Zetasizer Nano

Assay Bis tris Densitometry Hybrid Peptides Protein Sds page Uranyl acetate

Corresponding Organization :

Other organizations : University of California, San Diego, La Jolla Bioengineering Institute

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Characterization of VLPs
Determination of VLP concentration
Confirmation of hybrid QβS100A9 assembly and peptide display
Integrity of VLPs confirmed by TEM
Particle size confirmed by DLS

control variables

None explicitly mentioned

controls

Positive controls: None mentioned
Negative controls: None mentioned

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