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A frequency response function (FRF)-based indirect inverse sub-structuring method is developed for the analysis of the dynamic characteristics of a three-substructure coupled product transport system.A closed-form analytical solution to inverse sub-structuring analysis of multi-substructure coupled product transport system is derived.The proposed method is validated by a lumped mass-spring-damper model, the predicted coupling stiffness is compared with the direct computation, showing exact agreement.Then, the FRF tests of a physical prototype of multi-substructure coupled product transport system are performed to further check the accuracy of the suggested method.The method developed offers an approach to predict the unknown coupling stiffness from measured FRFs purely.The suggested method may help to obtain the main controlling factors and contributions from the various structure-borne paths for product transport system, which may certainly facilitate the cushioning packaging design.