One of the main surgical goals when performing a total knee replacement (TKR) is to ensure the implants are properly aligned and correctly sized; however, understanding the effect of alignment and rotation on the biomechanics of the knee during functional activities is limited. Cardiff University has unique access to a group of local patients who have relatively high frequency of poor alignment, and early failure. This provides a rare insight into how malalignment of TKR's can affect patients from a clinical and biomechanical point of view to determine how to best align a TKR. This study aims to explore relationship clinical surgical measurements of Implant alignment with in-vivo joint kinematics. 28 patient volunteers (with 32 Kinemax (Stryker) TKR's were recruited. Patients undertook single plane video fluoroscopy of the knee during a step-up and step-down task to determine TKR in-vivo kinematics and centre of rotation (COR). Joint Track image registration software (University of Florida, USA) was used to match CAD models of the implant to the x-ray images. Hip-Knee-Ankle (HKA) was measured using long-leg radiographs to determine frontal plane alignment. Posterior tibial slope angle was calculated using radiographs. An independent sample t-test was used to explore differences between neutral (HKA:-2° to 2°), varus (≥2°) and valgus alignment (≤-2°) groups. Other measures were explored across the whole cohort using Pearson's correlations (SPSS V23). There was found to be no statistical difference between groups or correlations for HKA. The exploratory analysis found that tibial slope correlated with Superior/Inferior translation ROM during step up (r=−0.601, p<0.001) and step down (r=−.512, p=0.03) the position of the COR heading towards the lateral (r=−.479, p=0.006) during step down. Initial results suggest no relationship between frontal plane alignment and in-vivo. Exploratory analyses have found other relationships that are worthy of further research and may be important in optimizing function.