Universities exert a considerable impact on the environment, economy, and society due to their complex infrastructures and multifaceted activities. This study applies the Life Cycle Sustainability Assessment (LCSA) methodology within the Sustainable Production Laboratory at the Federal University of São Carlos—Sorocaba campus, São Paulo, Brazil, to evaluate and enhance the sustainability performance of the laboratory. The LCSA framework integrates Environmental Life Cycle Assessment (LCA), Social Life Cycle Assessment (S-LCA), and Life Cycle Costing (LCC) to identify critical sustainability hotspots. The findings reveal that transportation constitutes the primary contributor to both environmental and social impacts, whereas expenditures related to human resources represent the predominant component of economic costs. To broaden the analytical scope, the study correlates LCSA outcomes with the Sustainable Development Goals (SDGs). The results underscore the necessity of implementing targeted strategies such as optimizing transportation systems, improving energy efficiency, and advancing resource management practices to mitigate adverse impacts. This research highlights the applicability and relevance of LCSA in higher education institutions, offering valuable insights for embedding sustainability principles into university operations. Furthermore, it contributes to the broader discourse on aligning institutional practices with global sustainable development agendas.