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This paper examines how managers struggled to implement the mandated separation of a unit from its parent organization within the English National Health Service (NHS) and how their attempts shaped subsequent inter-organizational dynamics. It demonstrates that enacting organizational separation, and thus aligning behavior with newly defined boundaries, involves potentially complex and interrelated forms of boundary work. Specifically, a qualitative analysis of this case suggests that attempts to separate stakeholders by limiting interaction across formal boundaries may unintentionally maintain connections and thus, paradoxically, counteract organizational separation. At the same time, attempts to establish connections may facilitate the organizational separation process by allowing stakeholders to negotiate and accept a new division of roles and responsibilities. Hence, I find that boundary spanning may be …