Understanding how innovation drives regional development has important economic and social outcomes. Models such as Smart Specialisation and the quadruple helix are increasingly adopted given their sensitivity to place-based contexts. But although innovation processes are dynamic and facilitated by individuals’ interactions, our understanding of how these connections are developed and sustained during helix collaboration development remains under-investigated. This research adopts an action research approach, focused on a regional Australian case study, to test a multi-disciplinary conceptual framework exploring key human-centred, micro-processes driving quadruple helix development: trust building; power relationships; regional readiness; and time and sphere centrality. The findings demonstrate the interactive nature of these processes, with trust-building facilitating the deployment of power bases by critical innovation agents which then foments regional readiness for change, subsequently driving the helix spheres towards overlap. These processes were also driven by changing sphere centrality and unique regional temporal structures. Practically, these outcomes offer insights into the human capital dynamics of regional helix collaborations, particularly for identifying the key individuals required to drive their development.