The examination of seated occupants’ ride comfort under whole-body vibration is a complex topic that involves multiple factors. Whole-body vibration refers to the mechanical vibration that is transmitted to the entire body through a supporting surface, such as a vehicle seat, when traveling on rough or uneven surfaces. There are several methods to assess ride comfort under whole-body vibration, such as subjective assessments, objective measurements, and mathematical models. Subjective assessments involve asking participants to rate their perceived level of discomfort or satisfaction during the vibration exposure, typically using a numerical scale or questionnaire. Objective measurements include accelerometers or vibration meters that record the actual physical vibrations transmitted to the body during the exposure. Mathematical models use various physiological and biomechanical parameters to predict the level of discomfort based on the vibration data. The examination of seated occupants ride comfort under whole-body vibration has been of great interest for many years. In this paper, a multi-body biomechanical model of a seated occupant with a backrest is proposed to perform ride comfort analysis. The novelty of the present model is that it represents complete passenger by including thighs, legs, and foot which were neglected in the past research. A multi-objective firefly algorithm is developed to evaluate the biomechanical parameters (mass, stiffness and damping) of the proposed model. Based on the optimized parameters, segmental transmissibilities are calculated and compared with experimental readings. The proposed model is then combined with a 7-dofs commercial car model to perform a ride comfort study. The ISO 2631-1:1997 ride comfort standards are used to compare the simulated segmental accelerations. Additionally, the influence of biomechanical parameters on most critical organs is analyzed to improve ride comfort. The outcomes of the analysis reveal that seated occupants perceive maximum vibration in the 3–6 Hz frequency range. To improve seated occupants' ride comfort, automotive designers must concentrate on the pelvis region. The adopted methodology and outcomes are helpful to evaluate protective measures in automobile industries. Furthermore, these procedures may be used to reduce the musculoskeletal disorders in seated occupants.