The WHO Unity Studies initiative engaged low- and middle-income countries in the implementation of standardized SARS-CoV-2 sero-epidemiological investigation protocols and timely sharing of comparable results for evidence-based action. To gain a deeper understanding of the methodological challenges faced when conducting seroprevalence studies in the Africa region, we conducted unstructured interviews with key study teams in five countries. We discuss the challenges identified: participant recruitment and retention, sample frame, sample and data management, data analysis and presentation to policy makers. Potential solutions to aid future implementation include preparedness actions such as the development of new tools, robust planning and practice.