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The layout of an office significantly affects human health, well-being and productivity in multi-occupancy offices, while the relationship and prioritization of various office layout factors have not been clarified. This study conducts a thorough investigation of office layout factors on occupant satisfaction. Existing pieces of literature were deconstructed and reorganized to extract five office layout factors. For a comparative investigation, an analytic hierarchy process-based online survey and a post-occupancy evaluation-based field survey focusing on academic buildings were implemented in China. The influence mechanisms in the satisfied and dissatisfied groups were explored together with the demographic differences by introducing the Kano model. The influence mechanism differences of the office layout factors between the priori and posteriori processes are emphasized. The results indicate that in academic buildings, the influence of a workstation location on satisfaction tends to be overestimated in the design stage, while the influence of space size tends to be underestimated in the design stage. Workstation orientation is a significant factor that is easily overlooked. Generally, an office layout design should focus more on ensuring the basic needs of all employees rather than creating design highlights. This study provides a new perspective to examine the gaps between actual satisfaction and expected satisfaction, and the obtained results can be directly used to guide user-centered office interior design.