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Science and technology-based agencies are increasingly subject to questions of social acceptance and credibility of their operations. Therefore, they engage in ‘legitimation practices’, which we define as the agencies’ own inside-out practices seeking to ensure that their activities enjoy broad acceptance and credibility among stakeholders. Sociological institutionalism suggests that as their formal mandate is comparable, agencies would develop similar practices through isomorphic processes. This paper compares the legitimation practices of three European agencies, EFSA, EMA and EPO through time. In particular, it examines the variation across agencies’ legitimation practices in terms of their institutionalized nature (how) and their issue-framings (what). The findings show that the agencies target their legitimacy practices to specific audiences, and rarely to the same general public, and consequently do not develop isomorphic features.