Background: The integration of health services with other sectors is hypothesised to support adaptation of health systems in response to coronavirus disease 2019 (COVID-19). This study identified barriers and enablers associated with intersectoral coordination at an early stage of the pandemic. The study focused on the roles played by the academic and private sector in different areas of public health planning and delivery concerning COVID-19 in Colombia. Methods: A qualitative approach was used to understand stakeholders’ experiences and perceptions of intersectoral working in response to COVID-19 in three Colombian cities (Bogotá, Cali and Cartagena). Between March and November 2020, data was collected via semi-structured interviews conducted online with 42 key actors, including representatives of governmental bodies, universities, and professional associations. The dataset was analysed thematically using a combination of inductive and deductive methods. Results: Organizations adjacent to the health system, including universities and the private sector, supported responses to COVID-19 by providing evidence to inform decision-making, additional service capacity, and supporting coordination (eg, convening intersectoral "roundtables"). The academic and private sector involvement in intersectoral coordination was stimulated by solidarity (being the "right thing to do") and motivation for supporting local companies (reopening the economy). Intersectoral working was influenced by pre-existing (substantive) and emerging (situational) enablers and barriers. Conclusion: This study showed that intersectoral coordination has played an important role in responding to COVID-19 in Colombia. Coordination was influenced by substantive and situational enablers and barriers. Based on our findings, policy-makers should focus on addressing substantive barriers to coordination, including the pre-existing tensions and mistrust among national and local healthcare actors, strict regulations and limited financial and human resources, while providing support for situational enablers, including alignment of public and private actors’ interests, intersectoral government support and establishing frequent communication channels and formal spaces of interaction among sector, in processes of decision-making.