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Africa reported lower COVID-19-related morbidity and mortality compared to other continents, despite widespread SARS-CoV-2 transmission and limited vaccine access. Proposed immunological explanations include potential pre-existing immunity such as cross-reactive humoral or cellular responses from earlier coronavirus exposures. However, functional immune responses to SARS-CoV-2 in African populations remain poorly characterized. To address this gap, we assessed post-pandemic neutralizing antibody responses against the SARS-CoV-2 D614G variant. We analyzed plasma samples from 989 participants in a cross-sectional survey in Ghana's Eastern and Greater Accra regions. A live virus neutralization assay using Vero E6 TMPRSS2 cells was employed to quantify SARS-CoV-2 D614G-specific neutralizing antibodies. Responses were assessed across collected demographic data. Urban participants exhibited higher median neutralizing antibody titers than rural counterparts, in both vaccinated and unvaccinated groups (p < 0.0001). Among unvaccinated individuals, median neutralizing antibody titers were comparable across age groups in urban settings. Vaccinated individuals showed elevated median titers across all age groups, with urban residents demonstrating stronger responses. Significant sex-based differences in neutralizing titres were also identified. Our findings reveal marked disparities in functional antibody responses between urban and rural populations, likely shaped by differences in SARS-CoV-2 exposure and vaccination. Continued surveillance and immunological profiling remain key for informing vaccine strategies and future pandemic preparedness.