Protocol for genome-scale differential flux analysis to interrogate metabolic differences from gene expression data

Deciphering the functional differences between diseased and healthy cells requires understanding the alterations in biochemical flux patterns. We present a genome-scale differential flux analysis (GS-DFA) protocol to elucidate these metabolic disparities by integrating condition-specific gene expression data into the human genome-scale metabolic model (humanGEM). In this protocol, we describe the steps to normalize and integrate data into the humanGEM and analyze differential flux across the biochemical network between diseased and healthy cells. For complete details on the use and execution of this protocol, please refer to Nanda et al.¹.