Background and Objectives: Healthcare workers are at heightened risk of SARS-CoV-2 infection. Understanding the duration and protective value of vaccine-induced immunity is critical to inform booster strategies. This study investigates longitudinal dynamics of anti-SARS-CoV-2 receptor-binding domain IgG (anti-RBD IgG) antibodies and their association with infection risk among vaccinated healthcare workers. Materials and Methods: A prospective cohort study was conducted at Vilnius University Hospital Santaros Klinikos, Lithuania. A total of 1778 healthcare workers who completed a primary COVID-19 vaccination series were followed. Blood samples were collected every three months to measure anti-RBD IgG levels. Participants also received up to three booster doses. COVID-19 was identified by PCR, antigen tests, or positive anti-nucleocapsid IgG. For serologically detected cases, infection timing was assigned to the interval between study visits. Antibody dynamics were analyzed across vaccination stages, time, age groups, and circulating SARS-CoV-2 variants. Results: Anti-RBD IgG titers peaked in the first quarter after primary vaccination (mean 7904 AU/mL), declined sharply by quarters 2–3, and rose substantially after booster doses. Following the first booster, titers increased to ~12,598 AU/mL in quarter 1 and continued rising through quarter 3. The highest levels were observed after the second booster (24,456 AU/mL in Q1), followed by gradual decline. A high-titer plateau persisted from quarters 6 to 9 (~21,000 AU/mL), followed by decline in quarters 10–11 and partial rebound in Q12. Approximately 49.6% of participants experienced COVID-19 during follow-up. Antibody response patterns were similar across age groups, with only minor transient differences. Conclusions: COVID-19 booster doses significantly enhance and prolong humoral immunity in healthcare workers compared with the primary vaccination series. However, antibody waning over time emphasizes the need for timely boosters, particularly during periods of variant circulation. These findings support continued booster vaccination and monitoring of long-term immune protection, although anti-RBD IgG should be interpreted as a surrogate marker of humoral rather than overall immunity.