Phosphoglycerate Kinase 1 (PGK1) is a crucial enzyme involved in glycolysis, and its abnormal expression has been implicated in the survival and proliferation of various cancers, making it a potential target for therapeutic intervention. In this study, BioAscent, in collaboration with the University of Edinburgh, undertook a comprehensive screening of BioAscent’s fragment library to identify novel small molecule binders to PGK1. Utilising a range of complementary biophysical techniques, including surface plasmon resonance (SPR) and thermal shift assays (TSA), the team successfully identified and characterised several fragment molecules that bind to PGK1. Using multiple biophysical methods ensured a robust and reliable identification process and showed the importance of using more than one technique when screening.