This article deals with Triple Helix (university, industry and government co-operation) from an institutional theory perspective. The empirical context is the Western Cape Region in South Africa and the focus is entrepreneurship development. The purpose is twofold: first, the existing Triple Helix model is adapted to the South African context; and second, facilities and impediments for working according to Triple Helix in South Africa are identified. The empirical material consists of a survey and three longitudinal case studies illustrating the degree of co-operation between the three parties. The article contributes to knowledge about how the Triple Helix model works on a regional level in a developing country. The study draws the following conclusions: when co-operation is to be identified between the three actors, only two of the three are involved; one missing link in the Triple Helix model is the focus on the entrepreneur; co-operation between the three parties are incidental rather than planned and there is lack of structure. In turn, some of these conclusions may be an effect of institutional changes on a national level. For a normative legacy, the article proposes a set of suggestions for incorporating all relevant parties on a practical level.