This study focuses on entrepreneurship in ecosystem-based adaptation to climate change (EbA). EbA uses ecosystem services and biodiversity to help society adapt to climate change. Several scholars argue that entrepreneurship, by introducing novel ideas and innovative solutions, can contribute to the planning and implementation of EbA. This study addresses a knowledge gap with regard to entrepreneurship in EbA by further elaborating the conceptual understanding of the role of entrepreneurs in EbA, the entrepreneurial opportunities that are developed in EbA and the interlinkages between entrepreneurs and opportunities. The main aim of this study is to better understand how entrepreneurs develop opportunities in EbA practice. It addresses this issue with a strong empirical focus, presenting several systematic within and between case study analyses. The results show that the creation and subsequent exploitation of opportunities in EbA by entrepreneurs is a dynamic and interactive process. To create opportunities in EbA, strategies are developed both individually and collectively. Also, entrepreneurs from the public and private sector and from civil society take over each other’s roles. A Qualitative Comparative Analysis (QCA) of 18 EbA cases show that two combinations of conditions explain successful exploitation of opportunities in EbA: facilitating policies and regulations with either high capital availability or with strong financial motives. Contrary to expectation, altruism is less important for successful exploitation. A follow-up frame analysis shows that six conditions for opportunity exploitation (i.e., the four conditions mentioned before as well as previous career experience and a social network) are constantly negotiated and shaped by the entrepreneurs and other actors involved. Aligning the temporal and spatial scale frames of entrepreneurs from the public and private sector appears especially challenging. Following the challenges encountered when calibrating data for the QCA study, this study further provides suggestions for more transparent data calibration, data presentation and sensitivity testing. This study provides a contribution to the governance of adaptation literature by increasing the conceptual understanding of entrepreneurship in adaptation. The study also provides insights in a method for systematic comparison of cases, potentially supporting concept development around climate change adaptation. Finally, the study makes a methodological contribution to the QCA literature and the more general literature on mixed methods.