Objective: This research intended to evaluate the antibody response to vaccination against hepatitis B during the first year of life. We also aimed to assess the association between the antibody response to hepatitis B vaccination and the polymorphism in HLA-G 14 bp. Material and methods: In this comparative cross-sectional study, 93 infants that received the hepatitis B vaccine at 2, 4, and 6 months old according to the World Health Organization immunization schedules, were evaluated for their antibodies to HBsAg (anti-HBs) following vaccination. Their genomic DNA was extracted and examined for the polymorphism in HLA-G 14 bp by PCR. Results: Among 93 vaccinated infants, 11 infants (11.8%) were non-responders to vaccination against hepatitis B (anti-HBs < 10 IU/mL). The mean antibody titers for responders and nonresponders were 77.65 ± 27.14 IU/mL and 4.45 ± 2.38 IU/mL, respectively. The 14-bp insertion allele was associated with an increased risk of failure o respond to hepatitis B vaccination (OR 3.72, 95% CI 1.32–10.6). Conclusion: This study reported, for the first time, that ins/ins genotype may be a risk factor for non-responsiveness to hepatitis B vaccination.