Previous studies have identified the importance of entrepreneurial and innovation ecosystems, however, few have considered in an integrated multi-level way, the key influencing factors on technology transfer mechanisms within these ecosystems. In particular, none have considered such factors from a technology transfer office (TTO) perspective. To address this gap, this research examines how macro–meso–micro factors shape the role of TTOs within an entrepreneurial and innovation ecosystem. Taking an in-depth exploratory approach set in the New Zealand context, data for this study was collected over two time periods, 4 years apart, and included inputs from all eight university TTO offices. Incorporating the perspective of TTO executives, our study unearths novel insights on a range of macro, meso and micro factors influencing their office, and how TTOs respond by synthesizing and balancing these factors across levels. The primary contribution of our research is extending the framing of TTO executives as typically portrayed in the literature, beyond important intermediaries between universities and industry, to pivotal cross-level brokers within entrepreneurial and innovation ecosystems.