The dynamics of transformations that the world is experiencing at global dimensions due to the intensity of technological changes demand sophisticated management tools to assess risks in the business and industrial sectors, aimed at ensuring investment security. The objective of this article is to analyse and propose the technical Due Diligence as a methodology to assess risks in Start-up ecosystems. The method used was mixed; a quantitative approach, and the qualitative approach, supported by a literature review with bibliographic arches. The sample was composed of thirty (30) experts, to whom a survey was applied, and to (10) of them, an interview that was subjected to a process of triangulation of the information, which was supported by documentary arches. The results showed the need to identify technological risks (product, service and process); commercial risks regarding the scalability of the business; and financial, legal, fiscal and environmental risks as part of a comprehensive and integral procedure.