Immunogenicity study of engineered ferritins with C- and N-terminus insertion of Epstein-Barr nuclear antigen 1 epitope

Immunogenicity and Vaccine Efficacy Boosted by Engineering Human Heavy Chain Ferritin and Chimeric Hepatitis B Virus Core Nanoparticles

“…The detailed calculation was shown in Supporting Information, S3. Another two control groups (n = 10/treatment) were injected with 200 μL of PBS (blank) and 100 μg of ovalbumin, 30 respectively. The serum was collected 10 days after the first (at day 24) and the second boost (at day 38) and stored at −80 °C before further measurement.…”

“…The detailed procedure followed our published research. 30 Short EBNA1 peptide (HPVGEADYFEY) was adsorbed for 14 h at 4 °C in 96-well plates. 30 After being blocked with the serum at 37 °C for 2 h, the plate was added with mouse sera of serial dilutions collected before (day 24 and day 38).…”

“…30 Short EBNA1 peptide (HPVGEADYFEY) was adsorbed for 14 h at 4 °C in 96-well plates. 30 After being blocked with the serum at 37 °C for 2 h, the plate was added with mouse sera of serial dilutions collected before (day 24 and day 38). The plate was further incubated at 37 °C for 2 h. After washing with PBS three times, 200 μL of goat anti-mouse IgG horseradish peroxidaseconjugated antibody of 1:5000 dilution was added to the plate and incubated at 37 °C for 2 h. 30 Color was developed by 3,3′,5,5′tetramethylbenzidine single-component substrate solution for 10 min.…”

“…Steps prior to plate washing were similar to the enzyme-linked immunosorbent assay (ELISA). 30 Antibodies goat anti-mouse IgG1 and IgG2a (1:1000 dilution, 100 μL) were loaded to each plate. 30 200 μL of rabbit anti-goat IgG horseradish peroxidase-conjugated antibody (1:5000 dilution) was loaded to the plate.…”

Biological Nanoparticles in Vaccine Development

“…Genetic modification of ferritin not only permits antigenic display, but also allows for the introduction of mutations that may improve functionality, such as addition of a cysteine group to the exterior to allow for conjugation of adjuvants via click chemistry ( Kamp et al, 2020 ). Due to its versatility, ferritin has been engineered as a vaccine platform to display antigens from various pathogens, including Borrelia burgdorferi ( Kamp et al, 2020 ), influenza ( Kanekiyo et al, 2013 ; Yassine et al, 2015 ; Qi et al, 2018 ; Wei et al, 2020 ), Epstein-Barr virus ( Kanekiyo et al, 2015 ; Qu et al, 2021b , 2021a ), hepatitis B ( Wang W. et al, 2020 ), rotavirus ( Li et al, 2019 ), and, most recently, SARS-Cov-2 ( Joyce et al, 2021 ; Kalathiya et al, 2021 ; Kang et al, 2021 ; King et al, 2021 ; Wuertz et al, 2021 ).…”