The development and application of in cellulo thermal proteome profiling assays in this study enabled a deeper investigation into the molecular mechanisms of the pathogenic NAA10 p.R83C variant. These findings offer novel insights into NAA10 function in the context of NAA10-related syndrome, a rare and currently understudied disorder. While the translational implications of this work are currently limited due to its exploratory nature, it lays a critical foundation for future research into the molecular pathology of this lethal disease. In addition, this work introduced a comparative analysis of various thermal proteome profiling assay formats, resulting in the proposal of a potentially novel high-throughput workflow. By incorporating additional replicates and datapoints, this approach aims to enhance both the resolution of protein stability modelling and the statistical robustness of the analysis. Although the methodology requires further refinement and validation, it presents a promising framework for future studies. Beyond its application to NAA10, this workflow may serve as a valuable tool for broader proteomic investigations across a range of disease models.