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The study aimed to assess the effect of five CpG ODNs in HBV Vaccine-immunized BALB/c mice and to identify the most effective CpG ODN adjuvant. In the vaccinated groups, those receiving supplementation with CpG 2395 exhibited a significant 4.4-fold elevation, resulting in a signal-to-noise ratio (S/N) value of 14.1 compared to vaccine group only (S/N = 3.22) by day 28 (p value < 0.0001). For mice immunized with the vaccine plus CpG ODNs, cytokine profiling using real-time quantitative polymerase chain reaction also revealed increased IL-6 expression levels and decreased TNF-α levels normalized with housekeeping gene GADPH. However, this increase in IL-6 was not statistically significant between the treated groups (P ≥ 0.3). Parameters for toxicity were within the normal range in all treatment groups. Statistical significance of differences was determined through One-way ANOVA and Student’s t-tests, with a P-value <0.05 being considered as statistically significant for all performed tests.